rneasy mini kit  (Qiagen)

 
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    Name:
    RNeasy Mini Kit
    Description:
    For purification of up to 100 µg total RNA from cells tissues and yeast Kit contents Qiagen RNeasy Mini Kit 50 preps 0 5 to 30mg Sample 30 to 100L Elution Volume Tissue Cells Sample Total RNA Purification Silica Technology Spin Column Format Ideal for Northern Dot and Slot Blotting End point RT PCR Quantitative Real time RT PCR For Purification of Up to 100g Total RNA from Cells Tissues and Yeast Includes 250 RNeasy Mini Spin Columns Collection Tubes 1 5mL and 2mL RNase free Reagents and Buffers Benefits Fast procedure delivering high quality total RNA in minutes Ready to use RNA for high performance in any downstream application Consistent RNA yields from small amounts of starting material No phenol chloroform extraction no CsCl gradients no LiCl or ethanol precipitatio
    Catalog Number:
    74104
    Price:
    314
    Category:
    RNeasy Mini Kit
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    Structured Review

    Qiagen rneasy mini kit
    RNeasy Mini Kit
    For purification of up to 100 µg total RNA from cells tissues and yeast Kit contents Qiagen RNeasy Mini Kit 50 preps 0 5 to 30mg Sample 30 to 100L Elution Volume Tissue Cells Sample Total RNA Purification Silica Technology Spin Column Format Ideal for Northern Dot and Slot Blotting End point RT PCR Quantitative Real time RT PCR For Purification of Up to 100g Total RNA from Cells Tissues and Yeast Includes 250 RNeasy Mini Spin Columns Collection Tubes 1 5mL and 2mL RNase free Reagents and Buffers Benefits Fast procedure delivering high quality total RNA in minutes Ready to use RNA for high performance in any downstream application Consistent RNA yields from small amounts of starting material No phenol chloroform extraction no CsCl gradients no LiCl or ethanol precipitatio
    https://www.bioz.com/result/rneasy mini kit/product/Qiagen
    Average 99 stars, based on 33750 article reviews
    Price from $9.99 to $1999.99
    rneasy mini kit - by Bioz Stars, 2020-07
    99/100 stars

    Images

    1) Product Images from "Optimizing exosomal RNA isolation for RNA-Seq analyses of archival sera specimens"

    Article Title: Optimizing exosomal RNA isolation for RNA-Seq analyses of archival sera specimens

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0196913

    Comparison of sera exosomal RNA using four different RNA extraction methods. (A) Total RNA yield from ultracentrifugation (UC) and ExoQuick (EQ) treated samples using the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and AllPrep DNA/RNA Mini Kit. (B) Demonstration of RNA quality measured by OD 260 /OD 280 in EQ and UC treated samples. Data are shown as the mean ± SD from six independent patient samples. *** P
    Figure Legend Snippet: Comparison of sera exosomal RNA using four different RNA extraction methods. (A) Total RNA yield from ultracentrifugation (UC) and ExoQuick (EQ) treated samples using the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and AllPrep DNA/RNA Mini Kit. (B) Demonstration of RNA quality measured by OD 260 /OD 280 in EQ and UC treated samples. Data are shown as the mean ± SD from six independent patient samples. *** P

    Techniques Used: RNA Extraction

    Exosomal RNAs are stable over two decades. Dot plots of total RNA yield and OD 260 /OD 280 from 105 EQ-treated archival patient samples using the RNeasy Mini Kit combined with TRIzol LS. Storage time for each sample is indicated on the x-axis. The Spearman’s rank correlations for RNA yield versus storing time and OD 260 /OD 280 versus storage time are 0.185 ( P = 0.06) and 0.04 ( P = 0.70), respectively.
    Figure Legend Snippet: Exosomal RNAs are stable over two decades. Dot plots of total RNA yield and OD 260 /OD 280 from 105 EQ-treated archival patient samples using the RNeasy Mini Kit combined with TRIzol LS. Storage time for each sample is indicated on the x-axis. The Spearman’s rank correlations for RNA yield versus storing time and OD 260 /OD 280 versus storage time are 0.185 ( P = 0.06) and 0.04 ( P = 0.70), respectively.

    Techniques Used:

    2) Product Images from "Monkeypox virus induces the synthesis of less dsRNA than vaccinia virus, and is more resistant to the anti-poxvirus drug, IBT, than vaccinia virus"

    Article Title: Monkeypox virus induces the synthesis of less dsRNA than vaccinia virus, and is more resistant to the anti-poxvirus drug, IBT, than vaccinia virus

    Journal: Virology

    doi: 10.1016/j.virol.2016.07.016

    Slot blot detection of dsRNA in extracts from virus-infected cells. Cells were infected with virus and then total RNA was extracted at multiple time points post-infection using the RNeasy Kit (Qiagen). Equal volumes of extract were then transferred onto
    Figure Legend Snippet: Slot blot detection of dsRNA in extracts from virus-infected cells. Cells were infected with virus and then total RNA was extracted at multiple time points post-infection using the RNeasy Kit (Qiagen). Equal volumes of extract were then transferred onto

    Techniques Used: Dot Blot, Infection

    3) Product Images from "Bias in recent miRBase annotations potentially associated with RNA quality issues"

    Article Title: Bias in recent miRBase annotations potentially associated with RNA quality issues

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-05070-0

    Experimental design. ( a ) Liver, heart and brain of male mice were harvested immediately after death, divided into 8 parts of about equal size, and stored at either 4 °C or at room temperature (RT) for the indicated time periods before RNA isolation. Experiments were performed in biological triplicates. RNA integrity was measured with Bioanalyzer. Gel-like image of brain tissue is given as example. MiRNA expression profiles of one replicate were measured using microarrays. ( b ) Liver tissue of 3 male mice was harvested immediately after death and divided into 5 parts of about equal size. Three parts were immediately transferred into RNAlater (0 h), two parts were stored for 96 h at room temperature (96 h). Two samples (0 h and 96 h) were isolated using standard procedure with miRNeasy Kit without DNase digestion. Two samples (0 h and 96 h) were isolated with optional DNase digestion to exclude DNA background. From the remaining undegraded sample (0 h), total RNA without small RNAs was isolated using RNeasy Kit with optional DNase digestion. Isolated RNA was further treated with 0 U, 0.026 U and 0.67 U RNase for 30 min to generate artificial RNA degradation. RNA integrity was measured with Bioanalyzer. MiRNA expression profiles of all replicates were measured using microarrays. The schematic drawings were prepared using the Biomedical-PPT-Toolkit-Suite from Motifolio Inc., USA.
    Figure Legend Snippet: Experimental design. ( a ) Liver, heart and brain of male mice were harvested immediately after death, divided into 8 parts of about equal size, and stored at either 4 °C or at room temperature (RT) for the indicated time periods before RNA isolation. Experiments were performed in biological triplicates. RNA integrity was measured with Bioanalyzer. Gel-like image of brain tissue is given as example. MiRNA expression profiles of one replicate were measured using microarrays. ( b ) Liver tissue of 3 male mice was harvested immediately after death and divided into 5 parts of about equal size. Three parts were immediately transferred into RNAlater (0 h), two parts were stored for 96 h at room temperature (96 h). Two samples (0 h and 96 h) were isolated using standard procedure with miRNeasy Kit without DNase digestion. Two samples (0 h and 96 h) were isolated with optional DNase digestion to exclude DNA background. From the remaining undegraded sample (0 h), total RNA without small RNAs was isolated using RNeasy Kit with optional DNase digestion. Isolated RNA was further treated with 0 U, 0.026 U and 0.67 U RNase for 30 min to generate artificial RNA degradation. RNA integrity was measured with Bioanalyzer. MiRNA expression profiles of all replicates were measured using microarrays. The schematic drawings were prepared using the Biomedical-PPT-Toolkit-Suite from Motifolio Inc., USA.

    Techniques Used: Mouse Assay, Isolation, Expressing

    4) Product Images from "Quantification of Small Non-Coding RNAs Allows an Accurate Comparison of miRNA Expression Profiles"

    Article Title: Quantification of Small Non-Coding RNAs Allows an Accurate Comparison of miRNA Expression Profiles

    Journal: Journal of Biomedicine and Biotechnology

    doi: 10.1155/2009/659028

    (a) Gel electrophoresis (polyacrylamide 15% stained with ethidium bromide) of LMW RNA samples (LCL) extracted with TRIzol reagent (lane 1), RNEasy kit (lane 2), and small RNA fraction enriched with MirVana kit (lane 3). LMW profile obtained with MirVana kit extraction is similar to that obtained with TRIzol reagent which is not shown for clarity. (b) Agilent 2100 Bioanalyzer electropherogram profile of LMW RNAs (LCL) extracted with TRIzol (black) superimposed on 5.8S (red), 5S (green), and tRNA (blue) bands eluted from polyacrylamide gel. (c) Lymphoblastoid (LCL) LMW RNA profile obtained after plotting the exported raw data from Agilent electropherogram ( ) together with the PeakFit fitted curve (solid line) and the component peak functions. Seven peaks below the LMW RNA profile were fitted by the software ( r 2 = 0.998693).
    Figure Legend Snippet: (a) Gel electrophoresis (polyacrylamide 15% stained with ethidium bromide) of LMW RNA samples (LCL) extracted with TRIzol reagent (lane 1), RNEasy kit (lane 2), and small RNA fraction enriched with MirVana kit (lane 3). LMW profile obtained with MirVana kit extraction is similar to that obtained with TRIzol reagent which is not shown for clarity. (b) Agilent 2100 Bioanalyzer electropherogram profile of LMW RNAs (LCL) extracted with TRIzol (black) superimposed on 5.8S (red), 5S (green), and tRNA (blue) bands eluted from polyacrylamide gel. (c) Lymphoblastoid (LCL) LMW RNA profile obtained after plotting the exported raw data from Agilent electropherogram ( ) together with the PeakFit fitted curve (solid line) and the component peak functions. Seven peaks below the LMW RNA profile were fitted by the software ( r 2 = 0.998693).

    Techniques Used: Nucleic Acid Electrophoresis, Staining, Software

    5) Product Images from "Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus"

    Article Title: Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus

    Journal: Brazilian Journal of Medical and Biological Research

    doi: 10.1590/1414-431X20154734

    Gel electrophoresis of total RNA that was obtained from sessile cells with sheared whole-cell lysis coupled to the RNeasy Mini Kit. Lanes 1 , 2 , and 3 represent three independent experiments using the methicillin-resistant S. aureus (MRSA) isolate BMB9393 ( A ) and the methicillin-susceptible S. aureus (MSSA) isolate HC474 ( B ).
    Figure Legend Snippet: Gel electrophoresis of total RNA that was obtained from sessile cells with sheared whole-cell lysis coupled to the RNeasy Mini Kit. Lanes 1 , 2 , and 3 represent three independent experiments using the methicillin-resistant S. aureus (MRSA) isolate BMB9393 ( A ) and the methicillin-susceptible S. aureus (MSSA) isolate HC474 ( B ).

    Techniques Used: Nucleic Acid Electrophoresis, Lysis

    6) Product Images from "Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus"

    Article Title: Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus

    Journal: Brazilian Journal of Medical and Biological Research

    doi: 10.1590/1414-431X20154734

    Gel electrophoresis of total RNA that was obtained from sessile cells with sheared whole-cell lysis coupled to the RNeasy Mini Kit. Lanes 1 , 2 , and 3 represent three independent experiments using the methicillin-resistant S. aureus (MRSA) isolate BMB9393 ( A ) and the methicillin-susceptible S. aureus (MSSA) isolate HC474 ( B ).
    Figure Legend Snippet: Gel electrophoresis of total RNA that was obtained from sessile cells with sheared whole-cell lysis coupled to the RNeasy Mini Kit. Lanes 1 , 2 , and 3 represent three independent experiments using the methicillin-resistant S. aureus (MRSA) isolate BMB9393 ( A ) and the methicillin-susceptible S. aureus (MSSA) isolate HC474 ( B ).

    Techniques Used: Nucleic Acid Electrophoresis, Lysis

    7) Product Images from "Amino Acids as Signaling Molecules Modulating Bone Turnover"

    Article Title: Amino Acids as Signaling Molecules Modulating Bone Turnover

    Journal: Bone

    doi: 10.1016/j.bone.2018.02.028

    Amino Acid Effects on BMSCs Panel A: (A) Effect of low amino acid levels on BMSC survival or of amino acid supplementation at a 1x (B) or 0.5x (C) level for three days followed by cell staining with Coomassie Blue. Higher magnification images are shown in the panels below. Essentially no BMSCs survived at the end of the three-day culture in M199 (A). Increasing concentrations of the amino acid mixture resulted in increased cell survival (B,C). Images representative of fifteen independent experiments. (D) Unstained cells grown in 1x amino-acid mixture. Panel B: RT-PCR for amino acid sensors: Total RNA was extracted from BMSC cells with the RNeasy mini kit (Qiagen). cDNA was synthesized using 1 μg total RNA (SuperScript III First-Strand Synthesis System, Invitrogen) and PCR performed as described in Methods. Panel C: RT-PCR for GPRC6A: To evaluate GPRC6A expression in bone cells we ran additional controls using the same conditions as in Panel B. As seen, we could detect GPRC6A in primary calvarial osteoblasts. Additional mouse tissues were used as positive and negative controls. OS= primary mouse calvarial osteoblasts; Hrt= mouse heart; Intest= mouse intestine; Kdn=mouse kidney, H 2 0 was a negative control
    Figure Legend Snippet: Amino Acid Effects on BMSCs Panel A: (A) Effect of low amino acid levels on BMSC survival or of amino acid supplementation at a 1x (B) or 0.5x (C) level for three days followed by cell staining with Coomassie Blue. Higher magnification images are shown in the panels below. Essentially no BMSCs survived at the end of the three-day culture in M199 (A). Increasing concentrations of the amino acid mixture resulted in increased cell survival (B,C). Images representative of fifteen independent experiments. (D) Unstained cells grown in 1x amino-acid mixture. Panel B: RT-PCR for amino acid sensors: Total RNA was extracted from BMSC cells with the RNeasy mini kit (Qiagen). cDNA was synthesized using 1 μg total RNA (SuperScript III First-Strand Synthesis System, Invitrogen) and PCR performed as described in Methods. Panel C: RT-PCR for GPRC6A: To evaluate GPRC6A expression in bone cells we ran additional controls using the same conditions as in Panel B. As seen, we could detect GPRC6A in primary calvarial osteoblasts. Additional mouse tissues were used as positive and negative controls. OS= primary mouse calvarial osteoblasts; Hrt= mouse heart; Intest= mouse intestine; Kdn=mouse kidney, H 2 0 was a negative control

    Techniques Used: Staining, Reverse Transcription Polymerase Chain Reaction, Synthesized, Polymerase Chain Reaction, Expressing, Negative Control

    8) Product Images from "PTK787/ZK22258 attenuates stellate cell activation and hepatic fibrosis in vivo by inhibiting VEGF signaling"

    Article Title: PTK787/ZK22258 attenuates stellate cell activation and hepatic fibrosis in vivo by inhibiting VEGF signaling

    Journal:

    doi: 10.1038/labinvest.2008.127

    mRNA expression of ( a ) procollagen, ( b ) TIMP-1, ( c ) MMP-9 and ( d ) CD31 in the livers of control and PTK/ZK-treated groups measured by real-time PCR. Total RNA was isolated from liver tissues using RNeasy Mini kit, and mRNA was quantitated by real-time
    Figure Legend Snippet: mRNA expression of ( a ) procollagen, ( b ) TIMP-1, ( c ) MMP-9 and ( d ) CD31 in the livers of control and PTK/ZK-treated groups measured by real-time PCR. Total RNA was isolated from liver tissues using RNeasy Mini kit, and mRNA was quantitated by real-time

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Isolation

    9) Product Images from "Quality Control of RNA Preservation and Extraction from Paraffin-Embedded Tissue: Implications for RT-PCR and Microarray Analysis"

    Article Title: Quality Control of RNA Preservation and Extraction from Paraffin-Embedded Tissue: Implications for RT-PCR and Microarray Analysis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070714

    Fixation has strong influence on RNA fragment size and impairs qRT-PCR results. Electropherograms show different RNA molecule size distribution in cryopreserved and FFPE/TFPE liver tissue samples. (A) In contrast to CRYO, ribosomal RNA peaks are absent and fragmentation of RNA has occurred in all fixed tissues. (B) RNA cleanup by Qiagen RNeasy Kit removes small (
    Figure Legend Snippet: Fixation has strong influence on RNA fragment size and impairs qRT-PCR results. Electropherograms show different RNA molecule size distribution in cryopreserved and FFPE/TFPE liver tissue samples. (A) In contrast to CRYO, ribosomal RNA peaks are absent and fragmentation of RNA has occurred in all fixed tissues. (B) RNA cleanup by Qiagen RNeasy Kit removes small (

    Techniques Used: Quantitative RT-PCR, Formalin-fixed Paraffin-Embedded

    10) Product Images from "A preliminary analysis of hepatitis C virus in pancreatic islet cells"

    Article Title: A preliminary analysis of hepatitis C virus in pancreatic islet cells

    Journal: Virology Journal

    doi: 10.1186/s12985-017-0905-3

    Total RNA from 3 different human islets donors was isolated using TRIzol reagent in combination with the RNeasy Mini kit followed by DNase treatment. 500 ng of total RNA were retrotranscribed using the Superscript III kit. The cDNAs obtained after retrotranscription were used as templates for quantitative real-time RT-PCR for mRNAs corresponding to the HCV entry factors CD81, occludin, claudin-1, and SR-B1. The relative amount of specific mRNA was normalized to Glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ). mRNA levels in Huh7.5 cells and human islets are relative to those in 293 T cells. Bars represent means ± SEM from three independent experiments
    Figure Legend Snippet: Total RNA from 3 different human islets donors was isolated using TRIzol reagent in combination with the RNeasy Mini kit followed by DNase treatment. 500 ng of total RNA were retrotranscribed using the Superscript III kit. The cDNAs obtained after retrotranscription were used as templates for quantitative real-time RT-PCR for mRNAs corresponding to the HCV entry factors CD81, occludin, claudin-1, and SR-B1. The relative amount of specific mRNA was normalized to Glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ). mRNA levels in Huh7.5 cells and human islets are relative to those in 293 T cells. Bars represent means ± SEM from three independent experiments

    Techniques Used: Isolation, Quantitative RT-PCR

    11) Product Images from "RNA extraction from self-assembling peptide hydrogels to allow qPCR analysis of encapsulated cells"

    Article Title: RNA extraction from self-assembling peptide hydrogels to allow qPCR analysis of encapsulated cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0197517

    A) Concentration of RNA extracted from HEK293 cells encapsulated in the four peptide hydrogels and cell-only controls using the three methods (see text for details). B Representative electrophoresis traces of total RNA extracted from cells encapsulated in the four peptide hydrogels and cell-only controls using the Tri Reagent and RNeasy MK methods and corresponding RIN values: cell-only controls (A+E), PGD-Alpha1 (B+F), PGD-Alpha2 (G), PGD-AlphaProB (C+H) and PGD-AlphaProC (D+I). C D) Representative UV spectra for RNA samples extracted using the TRI Reagent (C) and the RNeasy MK (D) methods from the four peptide hydrogels and cell-only control.
    Figure Legend Snippet: A) Concentration of RNA extracted from HEK293 cells encapsulated in the four peptide hydrogels and cell-only controls using the three methods (see text for details). B Representative electrophoresis traces of total RNA extracted from cells encapsulated in the four peptide hydrogels and cell-only controls using the Tri Reagent and RNeasy MK methods and corresponding RIN values: cell-only controls (A+E), PGD-Alpha1 (B+F), PGD-Alpha2 (G), PGD-AlphaProB (C+H) and PGD-AlphaProC (D+I). C D) Representative UV spectra for RNA samples extracted using the TRI Reagent (C) and the RNeasy MK (D) methods from the four peptide hydrogels and cell-only control.

    Techniques Used: Concentration Assay, Electrophoresis

    Ct values obtained for RT-qPCR performed using RNA extracted from the four peptide hydrogels as a template. RNA isolated from cells in suspension was used as a control. The RNA extracted using either the TRI Reagent method (A-B) or RNeasy MK method (C+D) was used as templates for the amplification of two housekeeping genes: GAPDH (A+C) and RPL13A (B+D). The cycle threshold (Ct) value was determined for three independent samples measured in triplicate (or two independent samples for PGD-Alpha1 using the RNeasy method). Data is shown as mean ± SEM. The mean values were compared to the non-encapsulated cell controls using a t-test; *, P ≤ 0.05.
    Figure Legend Snippet: Ct values obtained for RT-qPCR performed using RNA extracted from the four peptide hydrogels as a template. RNA isolated from cells in suspension was used as a control. The RNA extracted using either the TRI Reagent method (A-B) or RNeasy MK method (C+D) was used as templates for the amplification of two housekeeping genes: GAPDH (A+C) and RPL13A (B+D). The cycle threshold (Ct) value was determined for three independent samples measured in triplicate (or two independent samples for PGD-Alpha1 using the RNeasy method). Data is shown as mean ± SEM. The mean values were compared to the non-encapsulated cell controls using a t-test; *, P ≤ 0.05.

    Techniques Used: Quantitative RT-PCR, Isolation, Amplification

    12) Product Images from "Comparison of RNA extraction methods to augment the sensitivity for the differentiation of vesicular stomatitis virus Indiana1 and New Jersey"

    Article Title: Comparison of RNA extraction methods to augment the sensitivity for the differentiation of vesicular stomatitis virus Indiana1 and New Jersey

    Journal: Journal of Clinical Laboratory Analysis

    doi: 10.1002/jcla.20439

    Reverse transcription polymerase chain reaction amplification of RNA extracted from VSV‐IND and VSV‐NJ spiked bovine lymph nodes by Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lanes 2–6: IND and NJ RNA with NJ Primer set; lanes 7–11: IND and NJ RNA with IND Primer set; lane 12–16: IND and NJ RNA with both IND and NJ primer sets; lane 17: Negative extraction control; lane 18: Negative PCR control; lane 19: 100 bp marker.
    Figure Legend Snippet: Reverse transcription polymerase chain reaction amplification of RNA extracted from VSV‐IND and VSV‐NJ spiked bovine lymph nodes by Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lanes 2–6: IND and NJ RNA with NJ Primer set; lanes 7–11: IND and NJ RNA with IND Primer set; lane 12–16: IND and NJ RNA with both IND and NJ primer sets; lane 17: Negative extraction control; lane 18: Negative PCR control; lane 19: 100 bp marker.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Amplification, Marker, Polymerase Chain Reaction

    Multiplex RT‐PCR sensitivity and specificity assay for VSV‐IND and VSV‐NJ detection. RNA extracted from serial dilutions of both types of VSV with titers of 10 3.625 TCID 50 /25 µl and 10 4.375 TCID 50 /25 µl, for IND and NJ, respectively, using Qiagen RNeasy mini kit. Lanes 2–5: typical positive VSV‐IND reactions; lane 6: negative VSV‐IND reaction; lane 7–10: positive VSV‐NJ reactions; lane 11: negative VSV‐NJ reaction; lanes 12–15: VSV‐IND and VSV‐NJ positive reactions utilizing both primer sets; lane 16: negative for VSV‐IND NJ reaction; CE: negative extraction control; CP: negative PCR control; M: DNA marker.
    Figure Legend Snippet: Multiplex RT‐PCR sensitivity and specificity assay for VSV‐IND and VSV‐NJ detection. RNA extracted from serial dilutions of both types of VSV with titers of 10 3.625 TCID 50 /25 µl and 10 4.375 TCID 50 /25 µl, for IND and NJ, respectively, using Qiagen RNeasy mini kit. Lanes 2–5: typical positive VSV‐IND reactions; lane 6: negative VSV‐IND reaction; lane 7–10: positive VSV‐NJ reactions; lane 11: negative VSV‐NJ reaction; lanes 12–15: VSV‐IND and VSV‐NJ positive reactions utilizing both primer sets; lane 16: negative for VSV‐IND NJ reaction; CE: negative extraction control; CP: negative PCR control; M: DNA marker.

    Techniques Used: Multiplex Assay, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Marker

    Reverse transcription polymerase chain reaction amplification of RNA extracted from spiked BHK‐21 cell suspensions by using Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lane 2–6: IND and NJ RNA with IND Primer set; lane 7–11: IND and NJ RNA with NJ Primer set; lane 12: Negative extraction control (IND primer set); lane 13–17: IND and NJ RNA with both IND and NJ primer sets; lane 18: Negative extraction control (NJ primer set); lane 19: Negative extraction control (IND and NJ primer sets); lane 20: Negative PCR control.
    Figure Legend Snippet: Reverse transcription polymerase chain reaction amplification of RNA extracted from spiked BHK‐21 cell suspensions by using Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lane 2–6: IND and NJ RNA with IND Primer set; lane 7–11: IND and NJ RNA with NJ Primer set; lane 12: Negative extraction control (IND primer set); lane 13–17: IND and NJ RNA with both IND and NJ primer sets; lane 18: Negative extraction control (NJ primer set); lane 19: Negative extraction control (IND and NJ primer sets); lane 20: Negative PCR control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Amplification, Marker, Polymerase Chain Reaction

    13) Product Images from "An Analytically and Diagnostically Sensitive RNA Extraction and RT-qPCR Protocol for Peripheral Blood Mononuclear Cells"

    Article Title: An Analytically and Diagnostically Sensitive RNA Extraction and RT-qPCR Protocol for Peripheral Blood Mononuclear Cells

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2020.00402

    RNA extraction evaluation. (A) Bioanalyser analysis of RNA Integritry Number (RIN), and nanospectrophotometer analysis of yield and concentration were obtained using three commercially-available ex traction kits with (+) or without post-extraction RNA purification and concentration. (B) RT-qPCR analysis of IFN- γ, RPL13a, SDHA , and TBP expression normalized to cell number (copies/10 6 cells) or cDNA concentration (copies/μL). 1 × 10 6 PBMCs paired samples were cultured with complete media (white) or PMA/Iono (gray) for 6 h. RNA was extracted using the RNeasy® Mini (Mini) Kit, RNeasy® Micro (Micro) Kit (both QIAGEN), or MagMAX™ mirVana ™ (MagMAX) Total RNA Isolation Kit (Applied Biosystems), with concentration step performed using the RNeasy® MiniElute Cleanup Kit (QIAGEN). All samples were reverse transcribed with Superscript™ III (Invitrogen). Data were analyzed using a two-way ANOVA with post-hoc Bonferroni's multiple-comparisons test (* P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001). Biological replicate ( n = 4), triplicate RNA extractions, with single reverse transcription reactions per extraction were performed. Sample mean calculated from the mean of the technical RNA extractions which were in turn calculated from the mean of the technical triplicate qPCR reactions. Biological mean ± biological SEM are shown.
    Figure Legend Snippet: RNA extraction evaluation. (A) Bioanalyser analysis of RNA Integritry Number (RIN), and nanospectrophotometer analysis of yield and concentration were obtained using three commercially-available ex traction kits with (+) or without post-extraction RNA purification and concentration. (B) RT-qPCR analysis of IFN- γ, RPL13a, SDHA , and TBP expression normalized to cell number (copies/10 6 cells) or cDNA concentration (copies/μL). 1 × 10 6 PBMCs paired samples were cultured with complete media (white) or PMA/Iono (gray) for 6 h. RNA was extracted using the RNeasy® Mini (Mini) Kit, RNeasy® Micro (Micro) Kit (both QIAGEN), or MagMAX™ mirVana ™ (MagMAX) Total RNA Isolation Kit (Applied Biosystems), with concentration step performed using the RNeasy® MiniElute Cleanup Kit (QIAGEN). All samples were reverse transcribed with Superscript™ III (Invitrogen). Data were analyzed using a two-way ANOVA with post-hoc Bonferroni's multiple-comparisons test (* P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001). Biological replicate ( n = 4), triplicate RNA extractions, with single reverse transcription reactions per extraction were performed. Sample mean calculated from the mean of the technical RNA extractions which were in turn calculated from the mean of the technical triplicate qPCR reactions. Biological mean ± biological SEM are shown.

    Techniques Used: RNA Extraction, Concentration Assay, Purification, Quantitative RT-PCR, Expressing, Cell Culture, Isolation, Real-time Polymerase Chain Reaction

    Reverse transcription evaluation. Four reverse transcription kits were evalued for relative qPCR signal for (A) maximal RNA yield, or (B) maximal RNA concentration. When maximizing RNA yield, RNA was extracted with MagMAX™ mirVana ™ (MagMAX) Total RNA Isolation Kit (Applied Biosystems); when maximizing concentration, RNA was concentrated with RNeasy® MiniElute Cleanup Kit (QIAGEN). RNA was reverse transcribed with either Superscript™ III (SSIII), Superscript™ IV (SSIV) (both Invitrogen), iScript™ Advanced (iScript) (BioRad) or High-Capacity (HC) (ThermoFisher) reverse transcription kits. RNA was extracted from 1 × 10 6 PBMCs pared samples, cultured with complete media (white) or PMA/Iono (gray) for 6 h, then IFN- γ, RPL13a, SDHA , and TBP mRNA expression was quantified. Data were compared with a two-way ANOVA (* P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001 for post-hoc Bonferroni's multiple-comparisons test). Biological replicate ( n = 4), single RNA extractions, with triplicate reverse transcription reactions per extraction were performed. Sample mean calculated from the mean of the reverse transcription reactions calculated from the mean of the technical triplicate qPCR reactions. Biological mean ± biological SEM are shown.
    Figure Legend Snippet: Reverse transcription evaluation. Four reverse transcription kits were evalued for relative qPCR signal for (A) maximal RNA yield, or (B) maximal RNA concentration. When maximizing RNA yield, RNA was extracted with MagMAX™ mirVana ™ (MagMAX) Total RNA Isolation Kit (Applied Biosystems); when maximizing concentration, RNA was concentrated with RNeasy® MiniElute Cleanup Kit (QIAGEN). RNA was reverse transcribed with either Superscript™ III (SSIII), Superscript™ IV (SSIV) (both Invitrogen), iScript™ Advanced (iScript) (BioRad) or High-Capacity (HC) (ThermoFisher) reverse transcription kits. RNA was extracted from 1 × 10 6 PBMCs pared samples, cultured with complete media (white) or PMA/Iono (gray) for 6 h, then IFN- γ, RPL13a, SDHA , and TBP mRNA expression was quantified. Data were compared with a two-way ANOVA (* P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001 for post-hoc Bonferroni's multiple-comparisons test). Biological replicate ( n = 4), single RNA extractions, with triplicate reverse transcription reactions per extraction were performed. Sample mean calculated from the mean of the reverse transcription reactions calculated from the mean of the technical triplicate qPCR reactions. Biological mean ± biological SEM are shown.

    Techniques Used: Real-time Polymerase Chain Reaction, Concentration Assay, Isolation, Cell Culture, Expressing

    Assay analytical sensitivity. Relative RT-qPCR signal for IFN- γ, RPL13a, SDHA , and TBP mRNA expression from log 10 dilutions of unstimulated PBMCs when (A) maximizing RNA yield, or (B) maximizing RNA concentration. When maximizing RNA yield, RNA was extracted with MagMAX™ mirVana ™ (MagMAX) Total RNA Isolation Kit (Applied Biosystems); when maximizing concentration, RNA was concentrated with RNeasy® MiniElute Cleanup Kit (QIAGEN). All samples were reverse transcribed with Superscript™ IV (Invitrogen). mRNA expression was determined by absolute-quantitative RT-qPCR and gene copy number per reaction was normalized to log 10 copies per reaction. Biological replicate ( n = 3), single RNA extractions, with single reverse transcription reactions per extraction were performed. Sample mean calculated from the mean of the technical triplicate qPCR reactions. Biological mean ± biological SEM are shown.
    Figure Legend Snippet: Assay analytical sensitivity. Relative RT-qPCR signal for IFN- γ, RPL13a, SDHA , and TBP mRNA expression from log 10 dilutions of unstimulated PBMCs when (A) maximizing RNA yield, or (B) maximizing RNA concentration. When maximizing RNA yield, RNA was extracted with MagMAX™ mirVana ™ (MagMAX) Total RNA Isolation Kit (Applied Biosystems); when maximizing concentration, RNA was concentrated with RNeasy® MiniElute Cleanup Kit (QIAGEN). All samples were reverse transcribed with Superscript™ IV (Invitrogen). mRNA expression was determined by absolute-quantitative RT-qPCR and gene copy number per reaction was normalized to log 10 copies per reaction. Biological replicate ( n = 3), single RNA extractions, with single reverse transcription reactions per extraction were performed. Sample mean calculated from the mean of the technical triplicate qPCR reactions. Biological mean ± biological SEM are shown.

    Techniques Used: Quantitative RT-PCR, Expressing, Concentration Assay, Isolation, Real-time Polymerase Chain Reaction

    qPCR optimization. (A) Experimental workflow for qPCR optimization. (B) Effect of stimulation on mRNA expression of reference genes RPL13a, SDHA , and TBP . 1 × 10 6 PBMCs paired samples were cultured with complete media (white), or stimulated with PMA/Iono control (gray) for 0, 6, 12, 16, 24, or 48 h. RNA was extracted using the RNeasy® Mini (Mini) Kit, and reverse transcribed with Superscript™ III (Invitrogen). RNA expression was determined by absolute quantitative RT-qPCR wherein number of gene copies per reaction was quantified by standard curve and normalized to cell number. Data were compared with a two-way ANOVA with post-hoc Bonferroni's multiple-comparisons test (** P ≤ 0.01; *** P ≤ 0.001). Biological replicate ( n = 3) single RNA extractions with single reverse transcription reactions per extraction were performed. Sample mean calculated from technical triplicate qPCR. Biological mean ± biological SEM are shown.
    Figure Legend Snippet: qPCR optimization. (A) Experimental workflow for qPCR optimization. (B) Effect of stimulation on mRNA expression of reference genes RPL13a, SDHA , and TBP . 1 × 10 6 PBMCs paired samples were cultured with complete media (white), or stimulated with PMA/Iono control (gray) for 0, 6, 12, 16, 24, or 48 h. RNA was extracted using the RNeasy® Mini (Mini) Kit, and reverse transcribed with Superscript™ III (Invitrogen). RNA expression was determined by absolute quantitative RT-qPCR wherein number of gene copies per reaction was quantified by standard curve and normalized to cell number. Data were compared with a two-way ANOVA with post-hoc Bonferroni's multiple-comparisons test (** P ≤ 0.01; *** P ≤ 0.001). Biological replicate ( n = 3) single RNA extractions with single reverse transcription reactions per extraction were performed. Sample mean calculated from technical triplicate qPCR. Biological mean ± biological SEM are shown.

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing, Cell Culture, RNA Expression, Quantitative RT-PCR

    14) Product Images from "Comparison of RNA Isolation Methods From Insect Larvae"

    Article Title: Comparison of RNA Isolation Methods From Insect Larvae

    Journal: Journal of Insect Science

    doi: 10.1093/jisesa/ieu130

    Agilent 2100 Bioanalyser electropherogram graphs showing RNA extracted using the RNeasy Mini Kit, a CTAB-based protocol, TRIzol reagent and the SV Total RNA kit Markers can be seen at 25 nt, 18S RNA subunit at 2,000 nt, the 28S large subunit at 3,800 nt, and a combination of 5S, 5.8S, and tRNAs at ± 180 nt.
    Figure Legend Snippet: Agilent 2100 Bioanalyser electropherogram graphs showing RNA extracted using the RNeasy Mini Kit, a CTAB-based protocol, TRIzol reagent and the SV Total RNA kit Markers can be seen at 25 nt, 18S RNA subunit at 2,000 nt, the 28S large subunit at 3,800 nt, and a combination of 5S, 5.8S, and tRNAs at ± 180 nt.

    Techniques Used:

    Pseudo-gel image produced using an Agilent 2100 Bioanalyser, showing the results of RNA extracted from T. leucotreta using the RNeasy Mini Kit, a CTAB-based protocol, TRIzol reagent, and the SV Total RNA kit. The 18S RNA subunit is visible at 2,000 nt, the 28S large subunit at 3,800 nt, and a combination of 5S, 5.8S, and tRNAs at ± 180 nt.
    Figure Legend Snippet: Pseudo-gel image produced using an Agilent 2100 Bioanalyser, showing the results of RNA extracted from T. leucotreta using the RNeasy Mini Kit, a CTAB-based protocol, TRIzol reagent, and the SV Total RNA kit. The 18S RNA subunit is visible at 2,000 nt, the 28S large subunit at 3,800 nt, and a combination of 5S, 5.8S, and tRNAs at ± 180 nt.

    Techniques Used: Produced

    15) Product Images from "Optimizing exosomal RNA isolation for RNA-Seq analyses of archival sera specimens"

    Article Title: Optimizing exosomal RNA isolation for RNA-Seq analyses of archival sera specimens

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0196913

    Comparison of sera exosomal RNA using four different RNA extraction methods. (A) Total RNA yield from ultracentrifugation (UC) and ExoQuick (EQ) treated samples using the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and AllPrep DNA/RNA Mini Kit. (B) Demonstration of RNA quality measured by OD 260 /OD 280 in EQ and UC treated samples. Data are shown as the mean ± SD from six independent patient samples. *** P
    Figure Legend Snippet: Comparison of sera exosomal RNA using four different RNA extraction methods. (A) Total RNA yield from ultracentrifugation (UC) and ExoQuick (EQ) treated samples using the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and AllPrep DNA/RNA Mini Kit. (B) Demonstration of RNA quality measured by OD 260 /OD 280 in EQ and UC treated samples. Data are shown as the mean ± SD from six independent patient samples. *** P

    Techniques Used: RNA Extraction

    Exosomal RNAs are stable over two decades. Dot plots of total RNA yield and OD 260 /OD 280 from 105 EQ-treated archival patient samples using the RNeasy Mini Kit combined with TRIzol LS. Storage time for each sample is indicated on the x-axis. The Spearman’s rank correlations for RNA yield versus storing time and OD 260 /OD 280 versus storage time are 0.185 ( P = 0.06) and 0.04 ( P = 0.70), respectively.
    Figure Legend Snippet: Exosomal RNAs are stable over two decades. Dot plots of total RNA yield and OD 260 /OD 280 from 105 EQ-treated archival patient samples using the RNeasy Mini Kit combined with TRIzol LS. Storage time for each sample is indicated on the x-axis. The Spearman’s rank correlations for RNA yield versus storing time and OD 260 /OD 280 versus storage time are 0.185 ( P = 0.06) and 0.04 ( P = 0.70), respectively.

    Techniques Used:

    16) Product Images from "Effective methods for the inactivation of Francisella tularensis"

    Article Title: Effective methods for the inactivation of Francisella tularensis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0225177

    The viability of F . tularensis SCHU P9 after treatments of commercial products. Bacterial viability was evaluated after the treatment using Cell Lysis Buffer (Cell Signaling Technology) and the RLT buffer supplied by RNeasy mini kit (Qiagen Ltd.,). (A) Bacteria suspended in Cell Lysis Buffer and CDM (control) were incubated at 4°C for the indicated time. (B) Bacterial pellets after the centrifugation at 12,000 × g for 2 min at 4°C were suspended in RLT buffer alone, the mixture of RLT buffer and 70% ethanol, and CDM (control). The samples were incubated at room temperature for 10 min. All incubated samples were centrifuged at 12,000 × g for 2 min at 4°C and the pellets were suspended in CDM. the mean CFU ± SD of control and the treatment samples are shown. Statistical significance was determined by two-way ANOVA (A) and one-way ANOVA (B) with the post hoc test (*** p
    Figure Legend Snippet: The viability of F . tularensis SCHU P9 after treatments of commercial products. Bacterial viability was evaluated after the treatment using Cell Lysis Buffer (Cell Signaling Technology) and the RLT buffer supplied by RNeasy mini kit (Qiagen Ltd.,). (A) Bacteria suspended in Cell Lysis Buffer and CDM (control) were incubated at 4°C for the indicated time. (B) Bacterial pellets after the centrifugation at 12,000 × g for 2 min at 4°C were suspended in RLT buffer alone, the mixture of RLT buffer and 70% ethanol, and CDM (control). The samples were incubated at room temperature for 10 min. All incubated samples were centrifuged at 12,000 × g for 2 min at 4°C and the pellets were suspended in CDM. the mean CFU ± SD of control and the treatment samples are shown. Statistical significance was determined by two-way ANOVA (A) and one-way ANOVA (B) with the post hoc test (*** p

    Techniques Used: Lysis, Incubation, Centrifugation

    17) Product Images from "The circadian PER2 enhancer Nobiletin reverses the deleterious effects of midazolam in myocardial ischemia and reperfusion injury"

    Article Title: The circadian PER2 enhancer Nobiletin reverses the deleterious effects of midazolam in myocardial ischemia and reperfusion injury

    Journal: Current pharmaceutical design

    doi: 10.2174/1381612824666180924102530

    Studies of cardiac Per2 regulation following exposure of wildtype mice to anesthetics. Wildtype mice were exposed to a single dose of pentobarbital (70 mg/kg i.p.), fentanyl (1 mg/kg i.p.), isoflurane (1% inhaled), ketamine (200mg/kg i.p.), propofol (200 mg/kg i.p.), or midazolam (200 mg/kg i.p.). Two hours later cardiac Per2 mRNA expression levels were analyzed. In a subset of experiments murine endothelia, fibroblasts or cardiomyocytes were exposed to vehicle (NaCl 0.9%) or midazolam for 6 hours. Total RNA was isolated by Qiazol Reagent (Qiagen) and chloroform extraction in conjunction with the RNeasy Mini Kit (Qiagen), following the manufacturer’s instructions (SA-Biosciences, Qiagen). cDNA from mRNA was generated using iScript (Bio-Rad) and transcript levels were determined by real-time RT-PCR (iCycler; Bio-Rad Laboratories Inc.). ( A ) Overview and timeline of all in vivo studies. (I) Screening of different anesthetics for their effect on mouse heart Per2 mRNA levels. (II) Myocardial ischemia and reperfusion studies. (III) Reactive oxygen species (ROS) measurements following myocardial ischemia. ( B ) Mouse cardiac Per2 mRNA levels two hours after exposure to different anesthetics. ( C ) Per2 mRNA levels from murine cardiac endothelia after 6 h of midazolam (50μM) exposure. ( D ) Per2 mRNA levels from murine cardiac fibroblasts after 6 h of midazolam (50μM) exposure. ( E ) Per2 mRNA levels from isolated murine cardiomyocytes after 6 h of midazolam (50μM) exposure; (mean±SD, n=4–6; p
    Figure Legend Snippet: Studies of cardiac Per2 regulation following exposure of wildtype mice to anesthetics. Wildtype mice were exposed to a single dose of pentobarbital (70 mg/kg i.p.), fentanyl (1 mg/kg i.p.), isoflurane (1% inhaled), ketamine (200mg/kg i.p.), propofol (200 mg/kg i.p.), or midazolam (200 mg/kg i.p.). Two hours later cardiac Per2 mRNA expression levels were analyzed. In a subset of experiments murine endothelia, fibroblasts or cardiomyocytes were exposed to vehicle (NaCl 0.9%) or midazolam for 6 hours. Total RNA was isolated by Qiazol Reagent (Qiagen) and chloroform extraction in conjunction with the RNeasy Mini Kit (Qiagen), following the manufacturer’s instructions (SA-Biosciences, Qiagen). cDNA from mRNA was generated using iScript (Bio-Rad) and transcript levels were determined by real-time RT-PCR (iCycler; Bio-Rad Laboratories Inc.). ( A ) Overview and timeline of all in vivo studies. (I) Screening of different anesthetics for their effect on mouse heart Per2 mRNA levels. (II) Myocardial ischemia and reperfusion studies. (III) Reactive oxygen species (ROS) measurements following myocardial ischemia. ( B ) Mouse cardiac Per2 mRNA levels two hours after exposure to different anesthetics. ( C ) Per2 mRNA levels from murine cardiac endothelia after 6 h of midazolam (50μM) exposure. ( D ) Per2 mRNA levels from murine cardiac fibroblasts after 6 h of midazolam (50μM) exposure. ( E ) Per2 mRNA levels from isolated murine cardiomyocytes after 6 h of midazolam (50μM) exposure; (mean±SD, n=4–6; p

    Techniques Used: Mouse Assay, Expressing, Isolation, Generated, Quantitative RT-PCR, In Vivo

    18) Product Images from "Synergistic Cooperation between Methamphetamine and HIV-1 gsp120 through the P13K/Akt Pathway Induces IL-6 but not IL-8 Expression in Astrocytes"

    Article Title: Synergistic Cooperation between Methamphetamine and HIV-1 gsp120 through the P13K/Akt Pathway Induces IL-6 but not IL-8 Expression in Astrocytes

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0052060

    Methamphetamine-mediated induction of IL-6 and IL-8 does not involve the p38-MAPK or JNK-MAPK pathways. SVGA astrocytes were seeded at 8×10 5 cells in 6-well plate. The cells were treated once every day with 500 μM of MA for 3 days. Cells were treated with inhibitors alone or inhibitors and MA. Treatments with 10 μM of either SB203580 or SP600125 were started 1 hour prior to the MA treatment. The cells were harvested 24 hours after the last treatment of MA and total mRNA was isolated using an RNeasy mini kit. The RNA was analyzed using real time RT-PCR in order to quantify the expression levels of IL-6 (A) and IL-8 (B). Each experiment was repeated at least 3 times with triplicates of each treatment. The p-value ≤0.01 (**) was considered statistically significant when calculated with student's t-test.
    Figure Legend Snippet: Methamphetamine-mediated induction of IL-6 and IL-8 does not involve the p38-MAPK or JNK-MAPK pathways. SVGA astrocytes were seeded at 8×10 5 cells in 6-well plate. The cells were treated once every day with 500 μM of MA for 3 days. Cells were treated with inhibitors alone or inhibitors and MA. Treatments with 10 μM of either SB203580 or SP600125 were started 1 hour prior to the MA treatment. The cells were harvested 24 hours after the last treatment of MA and total mRNA was isolated using an RNeasy mini kit. The RNA was analyzed using real time RT-PCR in order to quantify the expression levels of IL-6 (A) and IL-8 (B). Each experiment was repeated at least 3 times with triplicates of each treatment. The p-value ≤0.01 (**) was considered statistically significant when calculated with student's t-test.

    Techniques Used: Isolation, Quantitative RT-PCR, Expressing

    19) Product Images from "DNA microarray-based experimental strategy for trustworthy expression profiling of the hippocampal genes by astaxanthin supplementation in adult mouse"

    Article Title: DNA microarray-based experimental strategy for trustworthy expression profiling of the hippocampal genes by astaxanthin supplementation in adult mouse

    Journal: Genomics Data

    doi: 10.1016/j.gdata.2015.11.001

    The experimental design for high-throughput DNA microarray analysis following astaxanthin (ASX) supplementation. (A) ASX supplementation was performed for four-weeks, and the mouse hippocampi was rapidly dissected and individually ground to a very fine powder in liquid nitrogen using pre-chilled mortar and pestle. (B) Total RNA was extracted from the right hippocampus using QIAGEN Rneasy Mini Kit, and their quality was confirmed both spectrometric method and formaldehyde-agarose gel electrophoresis. After this step, total RNA for each control and treatment (0.1% and 0.5% ASX) was pooled in each group ( n = 4). (C) The design of DNA microarray chip was followed as per the dye-swap labeling (CON × 0.1% ASX and CON × 0.5% ASX). The differentially expressed gene list was generated according to ≧/≦ 1.5/0.75-fold compared to CON. (D) As next step after the microarray, further confirmation of changed gene by RT-PCR, and detailed annotation of data sets and bioinformatics using Ingenuity Pathway Analysis (IPA) will be performed (manuscript to be published elsewhere).
    Figure Legend Snippet: The experimental design for high-throughput DNA microarray analysis following astaxanthin (ASX) supplementation. (A) ASX supplementation was performed for four-weeks, and the mouse hippocampi was rapidly dissected and individually ground to a very fine powder in liquid nitrogen using pre-chilled mortar and pestle. (B) Total RNA was extracted from the right hippocampus using QIAGEN Rneasy Mini Kit, and their quality was confirmed both spectrometric method and formaldehyde-agarose gel electrophoresis. After this step, total RNA for each control and treatment (0.1% and 0.5% ASX) was pooled in each group ( n = 4). (C) The design of DNA microarray chip was followed as per the dye-swap labeling (CON × 0.1% ASX and CON × 0.5% ASX). The differentially expressed gene list was generated according to ≧/≦ 1.5/0.75-fold compared to CON. (D) As next step after the microarray, further confirmation of changed gene by RT-PCR, and detailed annotation of data sets and bioinformatics using Ingenuity Pathway Analysis (IPA) will be performed (manuscript to be published elsewhere).

    Techniques Used: High Throughput Screening Assay, Microarray, Agarose Gel Electrophoresis, Chromatin Immunoprecipitation, Labeling, Generated, Reverse Transcription Polymerase Chain Reaction, Indirect Immunoperoxidase Assay

    20) Product Images from "Comparison of RNA extraction methods to augment the sensitivity for the differentiation of vesicular stomatitis virus Indiana1 and New Jersey"

    Article Title: Comparison of RNA extraction methods to augment the sensitivity for the differentiation of vesicular stomatitis virus Indiana1 and New Jersey

    Journal: Journal of Clinical Laboratory Analysis

    doi: 10.1002/jcla.20439

    Reverse transcription polymerase chain reaction amplification of RNA extracted from VSV‐IND and VSV‐NJ spiked bovine lymph nodes by Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lanes 2–6: IND and NJ RNA with NJ Primer set; lanes 7–11: IND and NJ RNA with IND Primer set; lane 12–16: IND and NJ RNA with both IND and NJ primer sets; lane 17: Negative extraction control; lane 18: Negative PCR control; lane 19: 100 bp marker.
    Figure Legend Snippet: Reverse transcription polymerase chain reaction amplification of RNA extracted from VSV‐IND and VSV‐NJ spiked bovine lymph nodes by Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lanes 2–6: IND and NJ RNA with NJ Primer set; lanes 7–11: IND and NJ RNA with IND Primer set; lane 12–16: IND and NJ RNA with both IND and NJ primer sets; lane 17: Negative extraction control; lane 18: Negative PCR control; lane 19: 100 bp marker.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Amplification, Marker, Polymerase Chain Reaction

    Multiplex RT‐PCR sensitivity and specificity assay for VSV‐IND and VSV‐NJ detection. RNA extracted from serial dilutions of both types of VSV with titers of 10 3.625 TCID 50 /25 µl and 10 4.375 TCID 50 /25 µl, for IND and NJ, respectively, using Qiagen RNeasy mini kit. Lanes 2–5: typical positive VSV‐IND reactions; lane 6: negative VSV‐IND reaction; lane 7–10: positive VSV‐NJ reactions; lane 11: negative VSV‐NJ reaction; lanes 12–15: VSV‐IND and VSV‐NJ positive reactions utilizing both primer sets; lane 16: negative for VSV‐IND NJ reaction; CE: negative extraction control; CP: negative PCR control; M: DNA marker.
    Figure Legend Snippet: Multiplex RT‐PCR sensitivity and specificity assay for VSV‐IND and VSV‐NJ detection. RNA extracted from serial dilutions of both types of VSV with titers of 10 3.625 TCID 50 /25 µl and 10 4.375 TCID 50 /25 µl, for IND and NJ, respectively, using Qiagen RNeasy mini kit. Lanes 2–5: typical positive VSV‐IND reactions; lane 6: negative VSV‐IND reaction; lane 7–10: positive VSV‐NJ reactions; lane 11: negative VSV‐NJ reaction; lanes 12–15: VSV‐IND and VSV‐NJ positive reactions utilizing both primer sets; lane 16: negative for VSV‐IND NJ reaction; CE: negative extraction control; CP: negative PCR control; M: DNA marker.

    Techniques Used: Multiplex Assay, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Marker

    Reverse transcription polymerase chain reaction amplification of RNA extracted from spiked BHK‐21 cell suspensions by using Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lane 2–6: IND and NJ RNA with IND Primer set; lane 7–11: IND and NJ RNA with NJ Primer set; lane 12: Negative extraction control (IND primer set); lane 13–17: IND and NJ RNA with both IND and NJ primer sets; lane 18: Negative extraction control (NJ primer set); lane 19: Negative extraction control (IND and NJ primer sets); lane 20: Negative PCR control.
    Figure Legend Snippet: Reverse transcription polymerase chain reaction amplification of RNA extracted from spiked BHK‐21 cell suspensions by using Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lane 2–6: IND and NJ RNA with IND Primer set; lane 7–11: IND and NJ RNA with NJ Primer set; lane 12: Negative extraction control (IND primer set); lane 13–17: IND and NJ RNA with both IND and NJ primer sets; lane 18: Negative extraction control (NJ primer set); lane 19: Negative extraction control (IND and NJ primer sets); lane 20: Negative PCR control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Amplification, Marker, Polymerase Chain Reaction

    21) Product Images from "Comparison of RNA extraction methods to augment the sensitivity for the differentiation of vesicular stomatitis virus Indiana1 and New Jersey"

    Article Title: Comparison of RNA extraction methods to augment the sensitivity for the differentiation of vesicular stomatitis virus Indiana1 and New Jersey

    Journal: Journal of Clinical Laboratory Analysis

    doi: 10.1002/jcla.20439

    Reverse transcription polymerase chain reaction amplification of RNA extracted from VSV‐IND and VSV‐NJ spiked bovine lymph nodes by Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lanes 2–6: IND and NJ RNA with NJ Primer set; lanes 7–11: IND and NJ RNA with IND Primer set; lane 12–16: IND and NJ RNA with both IND and NJ primer sets; lane 17: Negative extraction control; lane 18: Negative PCR control; lane 19: 100 bp marker.
    Figure Legend Snippet: Reverse transcription polymerase chain reaction amplification of RNA extracted from VSV‐IND and VSV‐NJ spiked bovine lymph nodes by Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lanes 2–6: IND and NJ RNA with NJ Primer set; lanes 7–11: IND and NJ RNA with IND Primer set; lane 12–16: IND and NJ RNA with both IND and NJ primer sets; lane 17: Negative extraction control; lane 18: Negative PCR control; lane 19: 100 bp marker.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Amplification, Marker, Polymerase Chain Reaction

    Multiplex RT‐PCR sensitivity and specificity assay for VSV‐IND and VSV‐NJ detection. RNA extracted from serial dilutions of both types of VSV with titers of 10 3.625 TCID 50 /25 µl and 10 4.375 TCID 50 /25 µl, for IND and NJ, respectively, using Qiagen RNeasy mini kit. Lanes 2–5: typical positive VSV‐IND reactions; lane 6: negative VSV‐IND reaction; lane 7–10: positive VSV‐NJ reactions; lane 11: negative VSV‐NJ reaction; lanes 12–15: VSV‐IND and VSV‐NJ positive reactions utilizing both primer sets; lane 16: negative for VSV‐IND NJ reaction; CE: negative extraction control; CP: negative PCR control; M: DNA marker.
    Figure Legend Snippet: Multiplex RT‐PCR sensitivity and specificity assay for VSV‐IND and VSV‐NJ detection. RNA extracted from serial dilutions of both types of VSV with titers of 10 3.625 TCID 50 /25 µl and 10 4.375 TCID 50 /25 µl, for IND and NJ, respectively, using Qiagen RNeasy mini kit. Lanes 2–5: typical positive VSV‐IND reactions; lane 6: negative VSV‐IND reaction; lane 7–10: positive VSV‐NJ reactions; lane 11: negative VSV‐NJ reaction; lanes 12–15: VSV‐IND and VSV‐NJ positive reactions utilizing both primer sets; lane 16: negative for VSV‐IND NJ reaction; CE: negative extraction control; CP: negative PCR control; M: DNA marker.

    Techniques Used: Multiplex Assay, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Marker

    Reverse transcription polymerase chain reaction amplification of RNA extracted from spiked BHK‐21 cell suspensions by using Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lane 2–6: IND and NJ RNA with IND Primer set; lane 7–11: IND and NJ RNA with NJ Primer set; lane 12: Negative extraction control (IND primer set); lane 13–17: IND and NJ RNA with both IND and NJ primer sets; lane 18: Negative extraction control (NJ primer set); lane 19: Negative extraction control (IND and NJ primer sets); lane 20: Negative PCR control.
    Figure Legend Snippet: Reverse transcription polymerase chain reaction amplification of RNA extracted from spiked BHK‐21 cell suspensions by using Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lane 2–6: IND and NJ RNA with IND Primer set; lane 7–11: IND and NJ RNA with NJ Primer set; lane 12: Negative extraction control (IND primer set); lane 13–17: IND and NJ RNA with both IND and NJ primer sets; lane 18: Negative extraction control (NJ primer set); lane 19: Negative extraction control (IND and NJ primer sets); lane 20: Negative PCR control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Amplification, Marker, Polymerase Chain Reaction

    22) Product Images from "Distinct roles for the kidney and systemic tissues in blood pressure regulation by the renin-angiotensin system"

    Article Title: Distinct roles for the kidney and systemic tissues in blood pressure regulation by the renin-angiotensin system

    Journal: Journal of Clinical Investigation

    doi: 10.1172/JCI200523378

    Kidney renin levels in the cross-transplantation groups. Kidneys were harvested at time of sacrifice. RNA was extracted using RNeasy protocol. Renin mRNA was quantitated by real-time PCR as described in Methods. *P
    Figure Legend Snippet: Kidney renin levels in the cross-transplantation groups. Kidneys were harvested at time of sacrifice. RNA was extracted using RNeasy protocol. Renin mRNA was quantitated by real-time PCR as described in Methods. *P

    Techniques Used: Transplantation Assay, Real-time Polymerase Chain Reaction

    23) Product Images from "New data on robustness of gene expression signatures in leukemia: comparison of three distinct total RNA preparation procedures"

    Article Title: New data on robustness of gene expression signatures in leukemia: comparison of three distinct total RNA preparation procedures

    Journal: BMC Genomics

    doi: 10.1186/1471-2164-8-188

    Study concept . (A) Total RNA of each of the first 24 samples had been extracted following three different total RNA purification methods A, B, and C. Method A: lysis of the mononuclear cells, followed by lysate homogenization (to reduce viscosity caused by high-molecular-weight cellular components and cell debris) using a biopolymer shredding system in a microcentrifuge spin-column format (QIAshredder, Qiagen) followed by total RNA purification (RNeasy Mini Kit, Qiagen). Method B: TRIzol RNA isolation (Invitrogen). Method C: TRIzol RNA isolation (Invitrogen) followed by an RNeasy purification step (RNeasy Mini Kit, Qiagen). The RNA purification step combines the selective binding properties of a silica-based membrane with the speed of microspin technology. It allows only RNA longer than 200 bases to bind to the silica membrane, providing an enriching for mRNA since nucleotides shorter than 200 nucleotides are selectively excluded. (B) For each of three additional samples, nine aliquots of mononuclear cells had been collected. Total RNA has been processed for each aliquot following one of the three methods and for each method three independent technical replicates were performed (A,A,A, B,B,B, C,C,C).
    Figure Legend Snippet: Study concept . (A) Total RNA of each of the first 24 samples had been extracted following three different total RNA purification methods A, B, and C. Method A: lysis of the mononuclear cells, followed by lysate homogenization (to reduce viscosity caused by high-molecular-weight cellular components and cell debris) using a biopolymer shredding system in a microcentrifuge spin-column format (QIAshredder, Qiagen) followed by total RNA purification (RNeasy Mini Kit, Qiagen). Method B: TRIzol RNA isolation (Invitrogen). Method C: TRIzol RNA isolation (Invitrogen) followed by an RNeasy purification step (RNeasy Mini Kit, Qiagen). The RNA purification step combines the selective binding properties of a silica-based membrane with the speed of microspin technology. It allows only RNA longer than 200 bases to bind to the silica membrane, providing an enriching for mRNA since nucleotides shorter than 200 nucleotides are selectively excluded. (B) For each of three additional samples, nine aliquots of mononuclear cells had been collected. Total RNA has been processed for each aliquot following one of the three methods and for each method three independent technical replicates were performed (A,A,A, B,B,B, C,C,C).

    Techniques Used: Purification, Lysis, Homogenization, Molecular Weight, Isolation, Binding Assay

    24) Product Images from "Changes in mRNA/protein expression and signaling pathways in in vivo passaged mouse ovarian cancer cells"

    Article Title: Changes in mRNA/protein expression and signaling pathways in in vivo passaged mouse ovarian cancer cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0197404

    Gene regulation confirmed by Q-PCR in mouse and human EOC cell lines. A-B. Mouse ID8 P0/P1 cells (A) or human EOC cells (B) were seeded into 6-well plates in attached or low-attached plates in suspension, RNAs were extracted with the RNeasy mini kit (Qiagen, Valencia, CA) and reverse transcribed by M-MLV reverse transcriptase. Quantitative real-time PCR was performed on a Light Cycler 480 (Roche, Indianapolis, IN) with a SYBR Green I Master Mix (Roche, Indianapolis, IN). mRNA abundance was normalized to GAPDH.
    Figure Legend Snippet: Gene regulation confirmed by Q-PCR in mouse and human EOC cell lines. A-B. Mouse ID8 P0/P1 cells (A) or human EOC cells (B) were seeded into 6-well plates in attached or low-attached plates in suspension, RNAs were extracted with the RNeasy mini kit (Qiagen, Valencia, CA) and reverse transcribed by M-MLV reverse transcriptase. Quantitative real-time PCR was performed on a Light Cycler 480 (Roche, Indianapolis, IN) with a SYBR Green I Master Mix (Roche, Indianapolis, IN). mRNA abundance was normalized to GAPDH.

    Techniques Used: Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, SYBR Green Assay

    25) Product Images from "Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus"

    Article Title: Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus

    Journal: Brazilian Journal of Medical and Biological Research

    doi: 10.1590/1414-431X20154734

    Gel electrophoresis of total RNA that was obtained from sessile cells with sheared whole-cell lysis coupled to the RNeasy Mini Kit. Lanes 1 , 2 , and 3 represent three independent experiments using the methicillin-resistant S. aureus (MRSA) isolate BMB9393 ( A ) and the methicillin-susceptible S. aureus (MSSA) isolate HC474 ( B ).
    Figure Legend Snippet: Gel electrophoresis of total RNA that was obtained from sessile cells with sheared whole-cell lysis coupled to the RNeasy Mini Kit. Lanes 1 , 2 , and 3 represent three independent experiments using the methicillin-resistant S. aureus (MRSA) isolate BMB9393 ( A ) and the methicillin-susceptible S. aureus (MSSA) isolate HC474 ( B ).

    Techniques Used: Nucleic Acid Electrophoresis, Lysis

    26) Product Images from "Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus"

    Article Title: Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus

    Journal: Brazilian Journal of Medical and Biological Research

    doi: 10.1590/1414-431X20154734

    Gel electrophoresis of total RNA that was obtained from sessile cells with sheared whole-cell lysis coupled to the RNeasy Mini Kit. Lanes 1 , 2 , and 3 represent three independent experiments using the methicillin-resistant S. aureus (MRSA) isolate BMB9393 ( A ) and the methicillin-susceptible S. aureus (MSSA) isolate HC474 ( B ).
    Figure Legend Snippet: Gel electrophoresis of total RNA that was obtained from sessile cells with sheared whole-cell lysis coupled to the RNeasy Mini Kit. Lanes 1 , 2 , and 3 represent three independent experiments using the methicillin-resistant S. aureus (MRSA) isolate BMB9393 ( A ) and the methicillin-susceptible S. aureus (MSSA) isolate HC474 ( B ).

    Techniques Used: Nucleic Acid Electrophoresis, Lysis

    27) Product Images from "Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus"

    Article Title: Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus

    Journal: Brazilian Journal of Medical and Biological Research

    doi: 10.1590/1414-431X20154734

    Gel electrophoresis of total RNA that was obtained from sessile cells with sheared whole-cell lysis coupled to the RNeasy Mini Kit. Lanes 1 , 2 , and 3 represent three independent experiments using the methicillin-resistant S. aureus (MRSA) isolate BMB9393 ( A ) and the methicillin-susceptible S. aureus (MSSA) isolate HC474 ( B ).
    Figure Legend Snippet: Gel electrophoresis of total RNA that was obtained from sessile cells with sheared whole-cell lysis coupled to the RNeasy Mini Kit. Lanes 1 , 2 , and 3 represent three independent experiments using the methicillin-resistant S. aureus (MRSA) isolate BMB9393 ( A ) and the methicillin-susceptible S. aureus (MSSA) isolate HC474 ( B ).

    Techniques Used: Nucleic Acid Electrophoresis, Lysis

    28) Product Images from "Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus"

    Article Title: Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus

    Journal: Brazilian Journal of Medical and Biological Research

    doi: 10.1590/1414-431X20154734

    Gel electrophoresis of total RNA that was obtained from sessile cells with sheared whole-cell lysis coupled to the RNeasy Mini Kit. Lanes 1 , 2 , and 3 represent three independent experiments using the methicillin-resistant S. aureus (MRSA) isolate BMB9393 ( A ) and the methicillin-susceptible S. aureus (MSSA) isolate HC474 ( B ).
    Figure Legend Snippet: Gel electrophoresis of total RNA that was obtained from sessile cells with sheared whole-cell lysis coupled to the RNeasy Mini Kit. Lanes 1 , 2 , and 3 represent three independent experiments using the methicillin-resistant S. aureus (MRSA) isolate BMB9393 ( A ) and the methicillin-susceptible S. aureus (MSSA) isolate HC474 ( B ).

    Techniques Used: Nucleic Acid Electrophoresis, Lysis

    29) Product Images from "HIV-1 gp120 induces type-1 programmed cell death through ER stress employing IRE1α, JNK and AP-1 pathway"

    Article Title: HIV-1 gp120 induces type-1 programmed cell death through ER stress employing IRE1α, JNK and AP-1 pathway

    Journal: Scientific Reports

    doi: 10.1038/srep18929

    HIV-1 gp120 increases XBP-1 splicing in time-dependent manner. ( A ) The schematic represents XBP-1 RNA processing. The loop represents the 26-nt region being removed during splicing of XBP-1 and theprimers were designed in order to amplify both the spliced and unspliced XBP-1. ( B ) SVGA cells were seeded at 2.5 × 10 5 cells/well in 12-well plates and transfected with pSyngp120 plasmid for 3, 6, 9 and 12 hours. The RNA were isolated using RNeasy mini kit and XBP-1 RNA were amplified using the primers shown in 3A. The amplified product was resolved using 3.5% agarose gel ( B ). The intensity of the spliced XBP-1 was measured and the mean ± S.E. was calculated from at least 3 experiments. The gel shown here is a representative of 3 independent experiments. The statistical significance was calculated using student’s t-test between 3H and respective time-points to show significant increase over time and the values were considered significant if p-value ≤ 0.01 (**).
    Figure Legend Snippet: HIV-1 gp120 increases XBP-1 splicing in time-dependent manner. ( A ) The schematic represents XBP-1 RNA processing. The loop represents the 26-nt region being removed during splicing of XBP-1 and theprimers were designed in order to amplify both the spliced and unspliced XBP-1. ( B ) SVGA cells were seeded at 2.5 × 10 5 cells/well in 12-well plates and transfected with pSyngp120 plasmid for 3, 6, 9 and 12 hours. The RNA were isolated using RNeasy mini kit and XBP-1 RNA were amplified using the primers shown in 3A. The amplified product was resolved using 3.5% agarose gel ( B ). The intensity of the spliced XBP-1 was measured and the mean ± S.E. was calculated from at least 3 experiments. The gel shown here is a representative of 3 independent experiments. The statistical significance was calculated using student’s t-test between 3H and respective time-points to show significant increase over time and the values were considered significant if p-value ≤ 0.01 (**).

    Techniques Used: Transfection, Plasmid Preparation, Isolation, Amplification, Agarose Gel Electrophoresis, IF-P

    30) Product Images from "Optimizing exosomal RNA isolation for RNA-Seq analyses of archival sera specimens"

    Article Title: Optimizing exosomal RNA isolation for RNA-Seq analyses of archival sera specimens

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0196913

    Comparison of sera exosomal RNA using four different RNA extraction methods. (A) Total RNA yield from ultracentrifugation (UC) and ExoQuick (EQ) treated samples using the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and AllPrep DNA/RNA Mini Kit. (B) Demonstration of RNA quality measured by OD 260 /OD 280 in EQ and UC treated samples. Data are shown as the mean ± SD from six independent patient samples. *** P
    Figure Legend Snippet: Comparison of sera exosomal RNA using four different RNA extraction methods. (A) Total RNA yield from ultracentrifugation (UC) and ExoQuick (EQ) treated samples using the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and AllPrep DNA/RNA Mini Kit. (B) Demonstration of RNA quality measured by OD 260 /OD 280 in EQ and UC treated samples. Data are shown as the mean ± SD from six independent patient samples. *** P

    Techniques Used: RNA Extraction

    Exosomal RNAs are stable over two decades. Dot plots of total RNA yield and OD 260 /OD 280 from 105 EQ-treated archival patient samples using the RNeasy Mini Kit combined with TRIzol LS. Storage time for each sample is indicated on the x-axis. The Spearman’s rank correlations for RNA yield versus storing time and OD 260 /OD 280 versus storage time are 0.185 ( P = 0.06) and 0.04 ( P = 0.70), respectively.
    Figure Legend Snippet: Exosomal RNAs are stable over two decades. Dot plots of total RNA yield and OD 260 /OD 280 from 105 EQ-treated archival patient samples using the RNeasy Mini Kit combined with TRIzol LS. Storage time for each sample is indicated on the x-axis. The Spearman’s rank correlations for RNA yield versus storing time and OD 260 /OD 280 versus storage time are 0.185 ( P = 0.06) and 0.04 ( P = 0.70), respectively.

    Techniques Used:

    Comparison of sera exosomal RNA using four different RNA extraction methods. (A) Total RNA yield from ultracentrifugation (UC) and ExoQuick (EQ) treated samples using the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and AllPrep DNA/RNA Mini Kit. (B) Demonstration of RNA quality measured by OD 260 /OD 280 in EQ and UC treated samples. Data are shown as the mean ± SD from six independent patient samples. *** P
    Figure Legend Snippet: Comparison of sera exosomal RNA using four different RNA extraction methods. (A) Total RNA yield from ultracentrifugation (UC) and ExoQuick (EQ) treated samples using the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and AllPrep DNA/RNA Mini Kit. (B) Demonstration of RNA quality measured by OD 260 /OD 280 in EQ and UC treated samples. Data are shown as the mean ± SD from six independent patient samples. *** P

    Techniques Used: RNA Extraction

    Exosomal RNAs are stable over two decades. Dot plots of total RNA yield and OD 260 /OD 280 from 105 EQ-treated archival patient samples using the RNeasy Mini Kit combined with TRIzol LS. Storage time for each sample is indicated on the x-axis. The Spearman’s rank correlations for RNA yield versus storing time and OD 260 /OD 280 versus storage time are 0.185 ( P = 0.06) and 0.04 ( P = 0.70), respectively.
    Figure Legend Snippet: Exosomal RNAs are stable over two decades. Dot plots of total RNA yield and OD 260 /OD 280 from 105 EQ-treated archival patient samples using the RNeasy Mini Kit combined with TRIzol LS. Storage time for each sample is indicated on the x-axis. The Spearman’s rank correlations for RNA yield versus storing time and OD 260 /OD 280 versus storage time are 0.185 ( P = 0.06) and 0.04 ( P = 0.70), respectively.

    Techniques Used:

    31) Product Images from "Optimizing exosomal RNA isolation for RNA-Seq analyses of archival sera specimens"

    Article Title: Optimizing exosomal RNA isolation for RNA-Seq analyses of archival sera specimens

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0196913

    Comparison of sera exosomal RNA using four different RNA extraction methods. (A) Total RNA yield from ultracentrifugation (UC) and ExoQuick (EQ) treated samples using the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and AllPrep DNA/RNA Mini Kit. (B) Demonstration of RNA quality measured by OD 260 /OD 280 in EQ and UC treated samples. Data are shown as the mean ± SD from six independent patient samples. *** P
    Figure Legend Snippet: Comparison of sera exosomal RNA using four different RNA extraction methods. (A) Total RNA yield from ultracentrifugation (UC) and ExoQuick (EQ) treated samples using the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and AllPrep DNA/RNA Mini Kit. (B) Demonstration of RNA quality measured by OD 260 /OD 280 in EQ and UC treated samples. Data are shown as the mean ± SD from six independent patient samples. *** P

    Techniques Used: RNA Extraction

    Exosomal RNAs are stable over two decades. Dot plots of total RNA yield and OD 260 /OD 280 from 105 EQ-treated archival patient samples using the RNeasy Mini Kit combined with TRIzol LS. Storage time for each sample is indicated on the x-axis. The Spearman’s rank correlations for RNA yield versus storing time and OD 260 /OD 280 versus storage time are 0.185 ( P = 0.06) and 0.04 ( P = 0.70), respectively.
    Figure Legend Snippet: Exosomal RNAs are stable over two decades. Dot plots of total RNA yield and OD 260 /OD 280 from 105 EQ-treated archival patient samples using the RNeasy Mini Kit combined with TRIzol LS. Storage time for each sample is indicated on the x-axis. The Spearman’s rank correlations for RNA yield versus storing time and OD 260 /OD 280 versus storage time are 0.185 ( P = 0.06) and 0.04 ( P = 0.70), respectively.

    Techniques Used:

    32) Product Images from "One-step RNA extraction for RT-qPCR detection of 2019-nCoV"

    Article Title: One-step RNA extraction for RT-qPCR detection of 2019-nCoV

    Journal: bioRxiv

    doi: 10.1101/2020.04.02.022384

    Side-by-side comparison of OP specimen from direct lysis in QE buffer vs. Qiagen RNeasy column purification. Ct values for probe sets N1, N2 and RNaseP shown using undilted and 1:10 dilutions of positive control counterparts at 100 copies/ul of plasmid DNA template and experimental OP specimen. A Ct value of 0 indicates no signal was detected.
    Figure Legend Snippet: Side-by-side comparison of OP specimen from direct lysis in QE buffer vs. Qiagen RNeasy column purification. Ct values for probe sets N1, N2 and RNaseP shown using undilted and 1:10 dilutions of positive control counterparts at 100 copies/ul of plasmid DNA template and experimental OP specimen. A Ct value of 0 indicates no signal was detected.

    Techniques Used: Lysis, Purification, Positive Control, Plasmid Preparation

    33) Product Images from "Interleukin-27 inhibition of HIV-1 involves an intermediate induction of type I interferon"

    Article Title: Interleukin-27 inhibition of HIV-1 involves an intermediate induction of type I interferon

    Journal:

    doi: 10.1182/blood-2009-03-211540

    IL-27 enhances macrophage APOBEC RNA expression . Monocyte-derived macrophages were treated 18 to 24 hours with IL-27 at 100 ng/mL or IFN-α at 10 ng/mL. Cells were lysed and processed for RNA following QIAGEN's RNeasy protocol, and gene transcription
    Figure Legend Snippet: IL-27 enhances macrophage APOBEC RNA expression . Monocyte-derived macrophages were treated 18 to 24 hours with IL-27 at 100 ng/mL or IFN-α at 10 ng/mL. Cells were lysed and processed for RNA following QIAGEN's RNeasy protocol, and gene transcription

    Techniques Used: RNA Expression, Derivative Assay

    34) Product Images from "Nucleic acids within urinary exosomes/microvesicles are potential biomarkers for renal disease"

    Article Title: Nucleic acids within urinary exosomes/microvesicles are potential biomarkers for renal disease

    Journal: Kidney international

    doi: 10.1038/ki.2010.106

    RNA extracted from whole urine cells and debris has a different RNA profile from that of tissue and urinary microvesicles ( a ) Analysis of RNA isolated from whole urine (exclusive of microvesicles that are not captured by the isolation technique) showed that a large yield of nucleic acids can be isolated (see the red profile). Processing of the isolated nucleic acids using the RNeasy Plus Micro kit (which removes gDNA) reveals that the majority of nucleic acids isolated using the ZR urine RNA isolation kit is DNA and the remaining RNA lacks rRNA peaks found in tissue and urinary exosomes. Red — nucleic acids isolated from whole urine without gDNA removal, blue — nucleic acids isolated from whole urine post gDNA removal using the RNeasy Plus Micro kit. ( b ) Isolation of microvesicles from the same urine sample revealed that the microvesicles retained a normal total RNA profile suggesting that RNA within whole cells may be less stable than that contained in urinary microvesicles. Red — without removal of gDNA, blue — sample processed using the RNeasy Plus Micro kit to remove contaminating gDNA. ( c ) Isolation of nucleic acids from the pellet formed during the 300 g spin revealed that the nucleic acid profile was different from that of microvesicles and that it contained a large amount of gDNA following processing using the RNeasy Plus Micro kit. Red — nucleic acids isolated from the 300 g pellet without gDNA removal, blue — nucleic acid isolated from the 300 g pellet post gDNA removal using the RNeasy Plus Micro kit. ( d ) Isolation of nucleic acids from pellets formed during the 17,000 g spin revealed that the nucleic acid profile was different to microvesicles and that it contained a large amount of gDNA following processing using the RNeasy Plus Micro kit. Red — nucleic acids isolated from the 17,000 g pellet without gDNA removal, blue — nucleic acids isolated from the 17,000 g pellet post gDNA removal using the RNeasy Plus Micro kit.
    Figure Legend Snippet: RNA extracted from whole urine cells and debris has a different RNA profile from that of tissue and urinary microvesicles ( a ) Analysis of RNA isolated from whole urine (exclusive of microvesicles that are not captured by the isolation technique) showed that a large yield of nucleic acids can be isolated (see the red profile). Processing of the isolated nucleic acids using the RNeasy Plus Micro kit (which removes gDNA) reveals that the majority of nucleic acids isolated using the ZR urine RNA isolation kit is DNA and the remaining RNA lacks rRNA peaks found in tissue and urinary exosomes. Red — nucleic acids isolated from whole urine without gDNA removal, blue — nucleic acids isolated from whole urine post gDNA removal using the RNeasy Plus Micro kit. ( b ) Isolation of microvesicles from the same urine sample revealed that the microvesicles retained a normal total RNA profile suggesting that RNA within whole cells may be less stable than that contained in urinary microvesicles. Red — without removal of gDNA, blue — sample processed using the RNeasy Plus Micro kit to remove contaminating gDNA. ( c ) Isolation of nucleic acids from the pellet formed during the 300 g spin revealed that the nucleic acid profile was different from that of microvesicles and that it contained a large amount of gDNA following processing using the RNeasy Plus Micro kit. Red — nucleic acids isolated from the 300 g pellet without gDNA removal, blue — nucleic acid isolated from the 300 g pellet post gDNA removal using the RNeasy Plus Micro kit. ( d ) Isolation of nucleic acids from pellets formed during the 17,000 g spin revealed that the nucleic acid profile was different to microvesicles and that it contained a large amount of gDNA following processing using the RNeasy Plus Micro kit. Red — nucleic acids isolated from the 17,000 g pellet without gDNA removal, blue — nucleic acids isolated from the 17,000 g pellet post gDNA removal using the RNeasy Plus Micro kit.

    Techniques Used: Isolation

    Analysis of nucleic acids associated with urinary microvesicles using the Agilent Bioanalyzer ( a ) Plot showing that microvesicles may co-isolate with extraneous DNA that can be removed by DNase digestion of the microvesicle pellet prior to lysis and nucleic acid extraction. Red — profile without DNase digestion, blue — profile with DNase digestion. ( b ) Plot showing that microvesicles do not co-isolate with detectable levels of extraneous RNA. Red — without RNase digestion, blue — with RNase digestion. ( c ) RNA isolated from rat kidney (red) and microvesicles (blue) exhibited a very similar profile, including the presence of 18S and 28S rRNA peaks. Both samples underwent processing using the RNeasy Plus Micro kit to remove genomic DNA (gDNA) contamination. ( d ) Urinary microvesicles contain a prominent ‘small RNA’ peak between 25–200 nt when miRNA isolation techniques are used. Red — kidney RNA isolated using RNeasy Plus Micro kit using the miRNA extraction method, blue — microvesicle RNA isolated with RNeasy Plus Micro kit using the miRNA extraction method. ( e ) Nucleic acids were isolated from microvesicles that had undergone RNase and DNase digestion on the outside before microvesicle lysis. During RNA extraction using the RNeasy Micro kit, half of the samples underwent on-column RNase digestion (see Materials and methods) while the other half underwent the same on-column incubation without the presence of RNase. Results revealed that RNase digestion was able to remove the majority of the profile, suggesting that RNA is the major nucleic acid within urinary microvesicles. Red — nucleic acid profile without intra-microvesicular RNase digestion, blue — nucleic acid profile with intra-microvesicular RNase digestion. ( f ) Further digestion with DNase following RNase digestion revealed that the remaining peak could be further reduced, suggesting that some material prone to DNase digestion remained in the sample potentially representing intra-exosomal DNA. Red — nucleic acid profile following intra-microvesicular on-column RNase digestion alone, blue — nucleic acid profile following both intra-microvesicular on-column RNase and DNase digestion. 18S and 28S rRNA peaks are indicated in ( a ). The peak at 25 nt represents an internal standard.
    Figure Legend Snippet: Analysis of nucleic acids associated with urinary microvesicles using the Agilent Bioanalyzer ( a ) Plot showing that microvesicles may co-isolate with extraneous DNA that can be removed by DNase digestion of the microvesicle pellet prior to lysis and nucleic acid extraction. Red — profile without DNase digestion, blue — profile with DNase digestion. ( b ) Plot showing that microvesicles do not co-isolate with detectable levels of extraneous RNA. Red — without RNase digestion, blue — with RNase digestion. ( c ) RNA isolated from rat kidney (red) and microvesicles (blue) exhibited a very similar profile, including the presence of 18S and 28S rRNA peaks. Both samples underwent processing using the RNeasy Plus Micro kit to remove genomic DNA (gDNA) contamination. ( d ) Urinary microvesicles contain a prominent ‘small RNA’ peak between 25–200 nt when miRNA isolation techniques are used. Red — kidney RNA isolated using RNeasy Plus Micro kit using the miRNA extraction method, blue — microvesicle RNA isolated with RNeasy Plus Micro kit using the miRNA extraction method. ( e ) Nucleic acids were isolated from microvesicles that had undergone RNase and DNase digestion on the outside before microvesicle lysis. During RNA extraction using the RNeasy Micro kit, half of the samples underwent on-column RNase digestion (see Materials and methods) while the other half underwent the same on-column incubation without the presence of RNase. Results revealed that RNase digestion was able to remove the majority of the profile, suggesting that RNA is the major nucleic acid within urinary microvesicles. Red — nucleic acid profile without intra-microvesicular RNase digestion, blue — nucleic acid profile with intra-microvesicular RNase digestion. ( f ) Further digestion with DNase following RNase digestion revealed that the remaining peak could be further reduced, suggesting that some material prone to DNase digestion remained in the sample potentially representing intra-exosomal DNA. Red — nucleic acid profile following intra-microvesicular on-column RNase digestion alone, blue — nucleic acid profile following both intra-microvesicular on-column RNase and DNase digestion. 18S and 28S rRNA peaks are indicated in ( a ). The peak at 25 nt represents an internal standard.

    Techniques Used: Lysis, Isolation, RNA Extraction, Incubation

    35) Product Images from "IL-22 modulates gut epithelial and immune barrier functions following acute alcohol exposure and burn injury"

    Article Title: IL-22 modulates gut epithelial and immune barrier functions following acute alcohol exposure and burn injury

    Journal: Shock (Augusta, Ga.)

    doi: 10.1097/SHK.0b013e3182749f96

    Decreased intestinal AMP expression one day post EtOH and burn injury Small intestine was harvested and equal weights of tissue (25 mg) from each experimental group were used for RNA extraction using the Qiagen RNeasy Mini Kit. Reg3β, Reg3γ and GAPDH were measured by qRT-PCR using specific probes. AMP expression was normalized to GAPDH expression via the ΔΔCT method. Values shown as mean + SEM, n=3–10 animals/group from two independent experiments. *, p
    Figure Legend Snippet: Decreased intestinal AMP expression one day post EtOH and burn injury Small intestine was harvested and equal weights of tissue (25 mg) from each experimental group were used for RNA extraction using the Qiagen RNeasy Mini Kit. Reg3β, Reg3γ and GAPDH were measured by qRT-PCR using specific probes. AMP expression was normalized to GAPDH expression via the ΔΔCT method. Values shown as mean + SEM, n=3–10 animals/group from two independent experiments. *, p

    Techniques Used: Expressing, RNA Extraction, Quantitative RT-PCR

    Treatment with IL-22 prevents the decrease in intestinal AMP expression one day post EtOH exposure and burn injury Day one after injury and IL-22 treatment, small intestine was harvested and equal weights of tissue (25 mg) from each experimental group were used for RNA extraction using the Qiagen RNeasy Mini Kit. Reg3β, Reg3γ and GAPDH were measured by qRT-PCR using specific probes. AMP expression was normalized to GAPDH expression via the ΔΔCT method. Values shown as mean + SEM, n=4–14 animals/group from three independent experiments. *, p
    Figure Legend Snippet: Treatment with IL-22 prevents the decrease in intestinal AMP expression one day post EtOH exposure and burn injury Day one after injury and IL-22 treatment, small intestine was harvested and equal weights of tissue (25 mg) from each experimental group were used for RNA extraction using the Qiagen RNeasy Mini Kit. Reg3β, Reg3γ and GAPDH were measured by qRT-PCR using specific probes. AMP expression was normalized to GAPDH expression via the ΔΔCT method. Values shown as mean + SEM, n=4–14 animals/group from three independent experiments. *, p

    Techniques Used: Expressing, RNA Extraction, Quantitative RT-PCR

    36) Product Images from "A Role for Phospholipase D3 in Myotube Formation"

    Article Title: A Role for Phospholipase D3 in Myotube Formation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0033341

    PLD3 expression increases during myogenic differentiation. ( A ) C2C12 cells were induced to differentiate for the indicated periods of time and RNA extracts prepared using RNeasy (Qiagen). Equal amounts of RNA were used as templates in quantitative RT-PCR with PLD3- and GAPDH-specific primers used for amplification, and performed in quadruplicate. The amount of PLD3 mRNA was normalized to the amounts amplified for GAPDH. ( B ) C2C12 cells were induced to differentiate for the indicated number of days and whole cell lysates collected. Equal amounts of lysate protein, as determined by Bradford reagent, were loaded onto SDS-polyacrylamide gels, and the subsequent blot probed with anti-PLD3 and anti-GAPDH antibodies. Representative blot of at least 3 independent experiments.
    Figure Legend Snippet: PLD3 expression increases during myogenic differentiation. ( A ) C2C12 cells were induced to differentiate for the indicated periods of time and RNA extracts prepared using RNeasy (Qiagen). Equal amounts of RNA were used as templates in quantitative RT-PCR with PLD3- and GAPDH-specific primers used for amplification, and performed in quadruplicate. The amount of PLD3 mRNA was normalized to the amounts amplified for GAPDH. ( B ) C2C12 cells were induced to differentiate for the indicated number of days and whole cell lysates collected. Equal amounts of lysate protein, as determined by Bradford reagent, were loaded onto SDS-polyacrylamide gels, and the subsequent blot probed with anti-PLD3 and anti-GAPDH antibodies. Representative blot of at least 3 independent experiments.

    Techniques Used: Expressing, Quantitative RT-PCR, Amplification

    37) Product Images from "Comparison of RNA extraction methods to augment the sensitivity for the differentiation of vesicular stomatitis virus Indiana1 and New Jersey"

    Article Title: Comparison of RNA extraction methods to augment the sensitivity for the differentiation of vesicular stomatitis virus Indiana1 and New Jersey

    Journal: Journal of Clinical Laboratory Analysis

    doi: 10.1002/jcla.20439

    Reverse transcription polymerase chain reaction amplification of RNA extracted from VSV‐IND and VSV‐NJ spiked bovine lymph nodes by Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lanes 2–6: IND and NJ RNA with NJ Primer set; lanes 7–11: IND and NJ RNA with IND Primer set; lane 12–16: IND and NJ RNA with both IND and NJ primer sets; lane 17: Negative extraction control; lane 18: Negative PCR control; lane 19: 100 bp marker.
    Figure Legend Snippet: Reverse transcription polymerase chain reaction amplification of RNA extracted from VSV‐IND and VSV‐NJ spiked bovine lymph nodes by Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lanes 2–6: IND and NJ RNA with NJ Primer set; lanes 7–11: IND and NJ RNA with IND Primer set; lane 12–16: IND and NJ RNA with both IND and NJ primer sets; lane 17: Negative extraction control; lane 18: Negative PCR control; lane 19: 100 bp marker.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Amplification, Marker, Polymerase Chain Reaction

    Multiplex RT‐PCR sensitivity and specificity assay for VSV‐IND and VSV‐NJ detection. RNA extracted from serial dilutions of both types of VSV with titers of 10 3.625 TCID 50 /25 µl and 10 4.375 TCID 50 /25 µl, for IND and NJ, respectively, using Qiagen RNeasy mini kit. Lanes 2–5: typical positive VSV‐IND reactions; lane 6: negative VSV‐IND reaction; lane 7–10: positive VSV‐NJ reactions; lane 11: negative VSV‐NJ reaction; lanes 12–15: VSV‐IND and VSV‐NJ positive reactions utilizing both primer sets; lane 16: negative for VSV‐IND NJ reaction; CE: negative extraction control; CP: negative PCR control; M: DNA marker.
    Figure Legend Snippet: Multiplex RT‐PCR sensitivity and specificity assay for VSV‐IND and VSV‐NJ detection. RNA extracted from serial dilutions of both types of VSV with titers of 10 3.625 TCID 50 /25 µl and 10 4.375 TCID 50 /25 µl, for IND and NJ, respectively, using Qiagen RNeasy mini kit. Lanes 2–5: typical positive VSV‐IND reactions; lane 6: negative VSV‐IND reaction; lane 7–10: positive VSV‐NJ reactions; lane 11: negative VSV‐NJ reaction; lanes 12–15: VSV‐IND and VSV‐NJ positive reactions utilizing both primer sets; lane 16: negative for VSV‐IND NJ reaction; CE: negative extraction control; CP: negative PCR control; M: DNA marker.

    Techniques Used: Multiplex Assay, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Marker

    Reverse transcription polymerase chain reaction amplification of RNA extracted from spiked BHK‐21 cell suspensions by using Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lane 2–6: IND and NJ RNA with IND Primer set; lane 7–11: IND and NJ RNA with NJ Primer set; lane 12: Negative extraction control (IND primer set); lane 13–17: IND and NJ RNA with both IND and NJ primer sets; lane 18: Negative extraction control (NJ primer set); lane 19: Negative extraction control (IND and NJ primer sets); lane 20: Negative PCR control.
    Figure Legend Snippet: Reverse transcription polymerase chain reaction amplification of RNA extracted from spiked BHK‐21 cell suspensions by using Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lane 2–6: IND and NJ RNA with IND Primer set; lane 7–11: IND and NJ RNA with NJ Primer set; lane 12: Negative extraction control (IND primer set); lane 13–17: IND and NJ RNA with both IND and NJ primer sets; lane 18: Negative extraction control (NJ primer set); lane 19: Negative extraction control (IND and NJ primer sets); lane 20: Negative PCR control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Amplification, Marker, Polymerase Chain Reaction

    38) Product Images from "Development of a low resource RNA extraction cassette based on surface tension valves"

    Article Title: Development of a low resource RNA extraction cassette based on surface tension valves

    Journal: ACS applied materials & interfaces

    doi: 10.1021/am2004009

    Comparison of RNA extracted from RSV infected (black bars) and uninfected (gray bars) HEp-2 cell lysates using the extraction cassette and RNeasy kit. Unextracted samples failed to report RSV RNA in either sample (mean ± s.d, n = 3).
    Figure Legend Snippet: Comparison of RNA extracted from RSV infected (black bars) and uninfected (gray bars) HEp-2 cell lysates using the extraction cassette and RNeasy kit. Unextracted samples failed to report RSV RNA in either sample (mean ± s.d, n = 3).

    Techniques Used: Infection

    The limit of detection of RNA detectable by RT-PCR after extraction from HEp-2 cell lysates spiked with known amounts of RSV RNA using either the continuous tubing extraction cassette (●) or the RNeasy kit (○) (mean ± s.d, n=3).
    Figure Legend Snippet: The limit of detection of RNA detectable by RT-PCR after extraction from HEp-2 cell lysates spiked with known amounts of RSV RNA using either the continuous tubing extraction cassette (●) or the RNeasy kit (○) (mean ± s.d, n=3).

    Techniques Used: Reverse Transcription Polymerase Chain Reaction

    The number of copies of RNA per μL extracted from RSV positive (black bars) and RSV negative (gray bars) nasal wash samples using five extraction methods were compared. Extractions were performed using prototype extraction cassette, RNeasy Mini
    Figure Legend Snippet: The number of copies of RNA per μL extracted from RSV positive (black bars) and RSV negative (gray bars) nasal wash samples using five extraction methods were compared. Extractions were performed using prototype extraction cassette, RNeasy Mini

    Techniques Used:

    Comparison of the percent of RSV RNA recovered after addition to TE buffer (A) or HEp-2 cell lysates (B) using the extraction cassette (left bars), RNeasy kit (middle bars), or no extraction (right bars) (mean ± s.d., n = 9). The recovery efficiency
    Figure Legend Snippet: Comparison of the percent of RSV RNA recovered after addition to TE buffer (A) or HEp-2 cell lysates (B) using the extraction cassette (left bars), RNeasy kit (middle bars), or no extraction (right bars) (mean ± s.d., n = 9). The recovery efficiency

    Techniques Used:

    39) Product Images from "Efficient extraction of small and large RNAs in bacteria for excellent total RNA sequencing and comprehensive transcriptome analysis"

    Article Title: Efficient extraction of small and large RNAs in bacteria for excellent total RNA sequencing and comprehensive transcriptome analysis

    Journal: BMC Research Notes

    doi: 10.1186/s13104-015-1726-3

    Electropherogram of RNA elutes obtained using three different RNA extraction methods. a RNA extracted from glucose-supplemented cells using TRI Reagent; b RNA extracted from PE-supplemented cells using TRI Reagent; c RNA extracted from glucose-supplemented cells using RNeasy kit; d RNA extracted from PE-supplemented cells using RNeasy kit; e RNA extracted from glucose-supplemented cells using Phenol-free kit; f RNA extracted from glucose-supplemented cells using Phenol-free kit. NA not available
    Figure Legend Snippet: Electropherogram of RNA elutes obtained using three different RNA extraction methods. a RNA extracted from glucose-supplemented cells using TRI Reagent; b RNA extracted from PE-supplemented cells using TRI Reagent; c RNA extracted from glucose-supplemented cells using RNeasy kit; d RNA extracted from PE-supplemented cells using RNeasy kit; e RNA extracted from glucose-supplemented cells using Phenol-free kit; f RNA extracted from glucose-supplemented cells using Phenol-free kit. NA not available

    Techniques Used: RNA Extraction

    40) Product Images from "Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus"

    Article Title: Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus

    Journal: Brazilian Journal of Medical and Biological Research

    doi: 10.1590/1414-431X20154734

    Gel electrophoresis of total RNA that was obtained from sessile cells with sheared whole-cell lysis coupled to the RNeasy Mini Kit. Lanes 1 , 2 , and 3 represent three independent experiments using the methicillin-resistant S. aureus (MRSA) isolate BMB9393 ( A ) and the methicillin-susceptible S. aureus (MSSA) isolate HC474 ( B ).
    Figure Legend Snippet: Gel electrophoresis of total RNA that was obtained from sessile cells with sheared whole-cell lysis coupled to the RNeasy Mini Kit. Lanes 1 , 2 , and 3 represent three independent experiments using the methicillin-resistant S. aureus (MRSA) isolate BMB9393 ( A ) and the methicillin-susceptible S. aureus (MSSA) isolate HC474 ( B ).

    Techniques Used: Nucleic Acid Electrophoresis, Lysis

    Related Articles

    RNA Extraction:

    Article Title: Quantification of Small Non-Coding RNAs Allows an Accurate Comparison of miRNA Expression Profiles
    Article Snippet: .. Total RNA Extraction and Small RNAs Enrichment Protocols Total RNA was extracted using three different methods: an acid phenol/guanidine isothiocyanate solution (TRIzol Reagent, Invitrogen), a glass-fiber filtration protocol (MirVana miRNA Isolation Kit, Ambion) that provides also a procedure to isolate and enrich low molecular weight (LMW) RNAs from higher molecular weight (HMW) RNAs, and another common glass-fiber purification protocol (RNEasy Mini Kit, Qiagen). ..

    Article Title: TRPM7‐mediated spontaneous Ca2+ entry regulates the proliferation and differentiation of human leukemia cell line K562. TRPM7‐mediated spontaneous Ca2+ entry regulates the proliferation and differentiation of human leukemia cell line K562
    Article Snippet: .. Reverse transcription polymerase reaction (RT‐PCR) and real‐time PCR Total RNA (about 1 μ g) was extracted from about a million of K562 cells with the total RNA extraction kit (RNeasy Minikit, Qiagen) and reverse transcribed (in a 10 μ L scale) by SuperScriptII (Thermo Fisher Scientific) with a randomized or an oligo‐dT primer to obtain the first strand DNA. .. PCR amplification (in a 20 μ L scale) was performed with a heat‐resistant DNA polymerase (PrimeSTAR GXL, Takara) in a thermal cycler (Biometra, Gottingen, Germany) according to the following protocol;denaturation at 94°C for 2 min followed by 17–35 cycles of: denaturation (94°C, 10s), annealing (60°C, 10s), and extension (72°C, 30s).

    Article Title: Astrocyte-elevated gene-1 confers resistance to pemetrexed in non-small cell lung cancer by upregulating thymidylate synthase expression
    Article Snippet: .. RNA extraction and quantitative real-time PCR Total RNA was extracted using a Qiagen mRNA easy mini kit (Qiagen). .. First-strand complementary DNA synthesis was performed from 2 μg of total RNA by using Invitrogen SuperScript Reverse Transcriptase (Thermo Fisher Scientific, Carlsbad, CA, USA).

    Filtration:

    Article Title: Quantification of Small Non-Coding RNAs Allows an Accurate Comparison of miRNA Expression Profiles
    Article Snippet: .. Total RNA Extraction and Small RNAs Enrichment Protocols Total RNA was extracted using three different methods: an acid phenol/guanidine isothiocyanate solution (TRIzol Reagent, Invitrogen), a glass-fiber filtration protocol (MirVana miRNA Isolation Kit, Ambion) that provides also a procedure to isolate and enrich low molecular weight (LMW) RNAs from higher molecular weight (HMW) RNAs, and another common glass-fiber purification protocol (RNEasy Mini Kit, Qiagen). ..

    Homogenization:

    Article Title: Monkeypox virus induces the synthesis of less dsRNA than vaccinia virus, and is more resistant to the anti-poxvirus drug, IBT, than vaccinia virus
    Article Snippet: .. Total RNA was extracted with the RNeasy Mini Kit, using QiaShredder homogenization and in-column DNase treatment as described by the manufacturer (Qiagen). .. An equal volume of total RNA (2 μL) was diluted in ddH2 0 (198 μL).

    Isolation:

    Article Title: Comparison of RNA Isolation Methods From Insect Larvae
    Article Snippet: .. Four methods including three commercial kits RNeasy Mini Kit (Qiagen), SV Total RNA isolation system (Promega), TRIzol reagent (Invitrogen), and a cetyl trimethylammonium bromide (CTAB)-based method were compared regarding their ability to isolate RNA from whole-body larvae ofThaumatotibia leucotreta (Meyrick),Thanatophilus micans (F.),Plutella xylostella (L.), andTenebrio molitor (L.). ..

    Article Title: Quantification of Small Non-Coding RNAs Allows an Accurate Comparison of miRNA Expression Profiles
    Article Snippet: .. Total RNA Extraction and Small RNAs Enrichment Protocols Total RNA was extracted using three different methods: an acid phenol/guanidine isothiocyanate solution (TRIzol Reagent, Invitrogen), a glass-fiber filtration protocol (MirVana miRNA Isolation Kit, Ambion) that provides also a procedure to isolate and enrich low molecular weight (LMW) RNAs from higher molecular weight (HMW) RNAs, and another common glass-fiber purification protocol (RNEasy Mini Kit, Qiagen). ..

    Article Title: Bias in recent miRBase annotations potentially associated with RNA quality issues
    Article Snippet: .. For determination of non-miRNA mediated background due to potential contamination with fragments of other RNAs during RNA degradation, total RNA was isolated from samples of three animals using RNeasy mini Kit (Qiagen, Hilden, Germany) with on column DNase digestion. .. We denominate this RNA as “RNA depleted of small RNAs”, being aware, that they are not 100% free of any miRNA or small RNA fractions, but the abundance of small RNAs in these samples is greatly reduced in comparison to the isolation procedure using the miRNeasy Kit.

    Article Title: Optimizing exosomal RNA isolation for RNA-Seq analyses of archival sera specimens
    Article Snippet: .. RNA was isolated from each ExoQuick prepared sample using the RNeasy Mini Kit combined with TRIzol LS. ..

    Purification:

    Article Title: Quantification of Small Non-Coding RNAs Allows an Accurate Comparison of miRNA Expression Profiles
    Article Snippet: .. Total RNA Extraction and Small RNAs Enrichment Protocols Total RNA was extracted using three different methods: an acid phenol/guanidine isothiocyanate solution (TRIzol Reagent, Invitrogen), a glass-fiber filtration protocol (MirVana miRNA Isolation Kit, Ambion) that provides also a procedure to isolate and enrich low molecular weight (LMW) RNAs from higher molecular weight (HMW) RNAs, and another common glass-fiber purification protocol (RNEasy Mini Kit, Qiagen). ..

    Real-time Polymerase Chain Reaction:

    Article Title: TRPM7‐mediated spontaneous Ca2+ entry regulates the proliferation and differentiation of human leukemia cell line K562. TRPM7‐mediated spontaneous Ca2+ entry regulates the proliferation and differentiation of human leukemia cell line K562
    Article Snippet: .. Reverse transcription polymerase reaction (RT‐PCR) and real‐time PCR Total RNA (about 1 μ g) was extracted from about a million of K562 cells with the total RNA extraction kit (RNeasy Minikit, Qiagen) and reverse transcribed (in a 10 μ L scale) by SuperScriptII (Thermo Fisher Scientific) with a randomized or an oligo‐dT primer to obtain the first strand DNA. .. PCR amplification (in a 20 μ L scale) was performed with a heat‐resistant DNA polymerase (PrimeSTAR GXL, Takara) in a thermal cycler (Biometra, Gottingen, Germany) according to the following protocol;denaturation at 94°C for 2 min followed by 17–35 cycles of: denaturation (94°C, 10s), annealing (60°C, 10s), and extension (72°C, 30s).

    Article Title: Astrocyte-elevated gene-1 confers resistance to pemetrexed in non-small cell lung cancer by upregulating thymidylate synthase expression
    Article Snippet: .. RNA extraction and quantitative real-time PCR Total RNA was extracted using a Qiagen mRNA easy mini kit (Qiagen). .. First-strand complementary DNA synthesis was performed from 2 μg of total RNA by using Invitrogen SuperScript Reverse Transcriptase (Thermo Fisher Scientific, Carlsbad, CA, USA).

    Concentration Assay:

    Article Title: Reprogramming Neutral Lipid Metabolism in Mouse Dendritic Leucocytes Hosting Live Leishmania amazonensis Amastigotes
    Article Snippet: .. DL extracted RNA integrity control Total RNA was extracted from MHC II+ DLs (RNeasy+ Mini-Kit, Qiagen) and its quality and concentration was determined using a NanoDrop ND-1000 micro-spectrophotometer (Kisker, http://www.kisker-biotech.com ) and an Agilent-2100 Bioanalyzer (Agilent, http://www.chem.agilent.com ). ..

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: TRPM7‐mediated spontaneous Ca2+ entry regulates the proliferation and differentiation of human leukemia cell line K562. TRPM7‐mediated spontaneous Ca2+ entry regulates the proliferation and differentiation of human leukemia cell line K562
    Article Snippet: .. Reverse transcription polymerase reaction (RT‐PCR) and real‐time PCR Total RNA (about 1 μ g) was extracted from about a million of K562 cells with the total RNA extraction kit (RNeasy Minikit, Qiagen) and reverse transcribed (in a 10 μ L scale) by SuperScriptII (Thermo Fisher Scientific) with a randomized or an oligo‐dT primer to obtain the first strand DNA. .. PCR amplification (in a 20 μ L scale) was performed with a heat‐resistant DNA polymerase (PrimeSTAR GXL, Takara) in a thermal cycler (Biometra, Gottingen, Germany) according to the following protocol;denaturation at 94°C for 2 min followed by 17–35 cycles of: denaturation (94°C, 10s), annealing (60°C, 10s), and extension (72°C, 30s).

    Molecular Weight:

    Article Title: Quantification of Small Non-Coding RNAs Allows an Accurate Comparison of miRNA Expression Profiles
    Article Snippet: .. Total RNA Extraction and Small RNAs Enrichment Protocols Total RNA was extracted using three different methods: an acid phenol/guanidine isothiocyanate solution (TRIzol Reagent, Invitrogen), a glass-fiber filtration protocol (MirVana miRNA Isolation Kit, Ambion) that provides also a procedure to isolate and enrich low molecular weight (LMW) RNAs from higher molecular weight (HMW) RNAs, and another common glass-fiber purification protocol (RNEasy Mini Kit, Qiagen). ..

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    Qiagen rneasy mini kit
    Reverse transcription polymerase chain reaction amplification of RNA extracted from VSV‐IND and VSV‐NJ spiked bovine lymph nodes by <t>Qiagen</t> <t>RNeasy</t> mini kit. Lane 1: 100 bp marker; lanes 2–6: IND and NJ RNA with NJ Primer set; lanes 7–11: IND and NJ RNA with IND Primer set; lane 12–16: IND and NJ RNA with both IND and NJ primer sets; lane 17: Negative extraction control; lane 18: Negative PCR control; lane 19: 100 bp marker.
    Rneasy Mini Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 33751 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Reverse transcription polymerase chain reaction amplification of RNA extracted from VSV‐IND and VSV‐NJ spiked bovine lymph nodes by Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lanes 2–6: IND and NJ RNA with NJ Primer set; lanes 7–11: IND and NJ RNA with IND Primer set; lane 12–16: IND and NJ RNA with both IND and NJ primer sets; lane 17: Negative extraction control; lane 18: Negative PCR control; lane 19: 100 bp marker.

    Journal: Journal of Clinical Laboratory Analysis

    Article Title: Comparison of RNA extraction methods to augment the sensitivity for the differentiation of vesicular stomatitis virus Indiana1 and New Jersey

    doi: 10.1002/jcla.20439

    Figure Lengend Snippet: Reverse transcription polymerase chain reaction amplification of RNA extracted from VSV‐IND and VSV‐NJ spiked bovine lymph nodes by Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lanes 2–6: IND and NJ RNA with NJ Primer set; lanes 7–11: IND and NJ RNA with IND Primer set; lane 12–16: IND and NJ RNA with both IND and NJ primer sets; lane 17: Negative extraction control; lane 18: Negative PCR control; lane 19: 100 bp marker.

    Article Snippet: RNA from various dilutions of each VSV type was extracted by using Qiagen RNeasy mini kit as per manufacturer's instructions (Qiagen, Valencia, CA).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Amplification, Marker, Polymerase Chain Reaction

    Multiplex RT‐PCR sensitivity and specificity assay for VSV‐IND and VSV‐NJ detection. RNA extracted from serial dilutions of both types of VSV with titers of 10 3.625 TCID 50 /25 µl and 10 4.375 TCID 50 /25 µl, for IND and NJ, respectively, using Qiagen RNeasy mini kit. Lanes 2–5: typical positive VSV‐IND reactions; lane 6: negative VSV‐IND reaction; lane 7–10: positive VSV‐NJ reactions; lane 11: negative VSV‐NJ reaction; lanes 12–15: VSV‐IND and VSV‐NJ positive reactions utilizing both primer sets; lane 16: negative for VSV‐IND NJ reaction; CE: negative extraction control; CP: negative PCR control; M: DNA marker.

    Journal: Journal of Clinical Laboratory Analysis

    Article Title: Comparison of RNA extraction methods to augment the sensitivity for the differentiation of vesicular stomatitis virus Indiana1 and New Jersey

    doi: 10.1002/jcla.20439

    Figure Lengend Snippet: Multiplex RT‐PCR sensitivity and specificity assay for VSV‐IND and VSV‐NJ detection. RNA extracted from serial dilutions of both types of VSV with titers of 10 3.625 TCID 50 /25 µl and 10 4.375 TCID 50 /25 µl, for IND and NJ, respectively, using Qiagen RNeasy mini kit. Lanes 2–5: typical positive VSV‐IND reactions; lane 6: negative VSV‐IND reaction; lane 7–10: positive VSV‐NJ reactions; lane 11: negative VSV‐NJ reaction; lanes 12–15: VSV‐IND and VSV‐NJ positive reactions utilizing both primer sets; lane 16: negative for VSV‐IND NJ reaction; CE: negative extraction control; CP: negative PCR control; M: DNA marker.

    Article Snippet: RNA from various dilutions of each VSV type was extracted by using Qiagen RNeasy mini kit as per manufacturer's instructions (Qiagen, Valencia, CA).

    Techniques: Multiplex Assay, Polymerase Chain Reaction, Marker

    Reverse transcription polymerase chain reaction amplification of RNA extracted from spiked BHK‐21 cell suspensions by using Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lane 2–6: IND and NJ RNA with IND Primer set; lane 7–11: IND and NJ RNA with NJ Primer set; lane 12: Negative extraction control (IND primer set); lane 13–17: IND and NJ RNA with both IND and NJ primer sets; lane 18: Negative extraction control (NJ primer set); lane 19: Negative extraction control (IND and NJ primer sets); lane 20: Negative PCR control.

    Journal: Journal of Clinical Laboratory Analysis

    Article Title: Comparison of RNA extraction methods to augment the sensitivity for the differentiation of vesicular stomatitis virus Indiana1 and New Jersey

    doi: 10.1002/jcla.20439

    Figure Lengend Snippet: Reverse transcription polymerase chain reaction amplification of RNA extracted from spiked BHK‐21 cell suspensions by using Qiagen RNeasy mini kit. Lane 1: 100 bp marker; lane 2–6: IND and NJ RNA with IND Primer set; lane 7–11: IND and NJ RNA with NJ Primer set; lane 12: Negative extraction control (IND primer set); lane 13–17: IND and NJ RNA with both IND and NJ primer sets; lane 18: Negative extraction control (NJ primer set); lane 19: Negative extraction control (IND and NJ primer sets); lane 20: Negative PCR control.

    Article Snippet: RNA from various dilutions of each VSV type was extracted by using Qiagen RNeasy mini kit as per manufacturer's instructions (Qiagen, Valencia, CA).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Amplification, Marker, Polymerase Chain Reaction

    Comparison of sera exosomal RNA using four different RNA extraction methods. (A) Total RNA yield from ultracentrifugation (UC) and ExoQuick (EQ) treated samples using the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and AllPrep DNA/RNA Mini Kit. (B) Demonstration of RNA quality measured by OD 260 /OD 280 in EQ and UC treated samples. Data are shown as the mean ± SD from six independent patient samples. *** P

    Journal: PLoS ONE

    Article Title: Optimizing exosomal RNA isolation for RNA-Seq analyses of archival sera specimens

    doi: 10.1371/journal.pone.0196913

    Figure Lengend Snippet: Comparison of sera exosomal RNA using four different RNA extraction methods. (A) Total RNA yield from ultracentrifugation (UC) and ExoQuick (EQ) treated samples using the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and AllPrep DNA/RNA Mini Kit. (B) Demonstration of RNA quality measured by OD 260 /OD 280 in EQ and UC treated samples. Data are shown as the mean ± SD from six independent patient samples. *** P

    Article Snippet: RNA was isolated from each ExoQuick prepared sample using the RNeasy Mini Kit combined with TRIzol LS.

    Techniques: RNA Extraction

    Exosomal RNAs are stable over two decades. Dot plots of total RNA yield and OD 260 /OD 280 from 105 EQ-treated archival patient samples using the RNeasy Mini Kit combined with TRIzol LS. Storage time for each sample is indicated on the x-axis. The Spearman’s rank correlations for RNA yield versus storing time and OD 260 /OD 280 versus storage time are 0.185 ( P = 0.06) and 0.04 ( P = 0.70), respectively.

    Journal: PLoS ONE

    Article Title: Optimizing exosomal RNA isolation for RNA-Seq analyses of archival sera specimens

    doi: 10.1371/journal.pone.0196913

    Figure Lengend Snippet: Exosomal RNAs are stable over two decades. Dot plots of total RNA yield and OD 260 /OD 280 from 105 EQ-treated archival patient samples using the RNeasy Mini Kit combined with TRIzol LS. Storage time for each sample is indicated on the x-axis. The Spearman’s rank correlations for RNA yield versus storing time and OD 260 /OD 280 versus storage time are 0.185 ( P = 0.06) and 0.04 ( P = 0.70), respectively.

    Article Snippet: RNA was isolated from each ExoQuick prepared sample using the RNeasy Mini Kit combined with TRIzol LS.

    Techniques:

    FCM detection of lipid bodies (LBs) in control C57BL/6 DLs and C57BL/6 DLs hosting Ds Red2 L. am amastigotes. Panel A. Representative analysis of the fluorescence of BODIPY 493/503 in MHC II + DLs. A1, A2. Control C57BL/6 DL cultures –i.e. not exposed to L. am amastigotes- were incubated or not at 34°C with 200 µM oleate for 21 hours. A3, A4. Ds Red2- L. am were added to DL cultures at a ratio of 5 Ds Red2- L. am per DL (+ L. am ). Oleic acid (200 µM) was added 3 hours post inoculation, (A4: +Oleate) or not (A3: −Oleate) for a further 21 hours at 34°C. Control and L. am -loaded DLs were then detached. LBs were stained with BODIPY 493/503 2 µg/ml in PBS for 30 minutes at 34°C. The cells were then incubated with anti MHC II- PE-Cy5 mAb to analyze only the MHC class II-positive DLs. FCM histograms show BODIPY 493/503 fluorescence for Ctrl (A1, A2: white histograms) and Ds Red2 L. am -hosting DLs (A3, A4) ( Ds Red2 + , black histograms) and amastigotes-free DLs ( Ds Red2 − , grey histograms). Mean fluorescence intensity was indicated for each histogram. Panel B: Statistical analysis of the BODIPY 493/503 MFI values. Mean BODIPY 493/503 fluorescence by MHC II + DLs was determined for n = 7 independent experiments. MFI in L. am -loaded cultures is shown for Ds Red2 + and Ds Red2 − DLs as black and grey bars, respectively. Statistical analyses were performed by the Mann Whitney test.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Reprogramming Neutral Lipid Metabolism in Mouse Dendritic Leucocytes Hosting Live Leishmania amazonensis Amastigotes

    doi: 10.1371/journal.pntd.0002276

    Figure Lengend Snippet: FCM detection of lipid bodies (LBs) in control C57BL/6 DLs and C57BL/6 DLs hosting Ds Red2 L. am amastigotes. Panel A. Representative analysis of the fluorescence of BODIPY 493/503 in MHC II + DLs. A1, A2. Control C57BL/6 DL cultures –i.e. not exposed to L. am amastigotes- were incubated or not at 34°C with 200 µM oleate for 21 hours. A3, A4. Ds Red2- L. am were added to DL cultures at a ratio of 5 Ds Red2- L. am per DL (+ L. am ). Oleic acid (200 µM) was added 3 hours post inoculation, (A4: +Oleate) or not (A3: −Oleate) for a further 21 hours at 34°C. Control and L. am -loaded DLs were then detached. LBs were stained with BODIPY 493/503 2 µg/ml in PBS for 30 minutes at 34°C. The cells were then incubated with anti MHC II- PE-Cy5 mAb to analyze only the MHC class II-positive DLs. FCM histograms show BODIPY 493/503 fluorescence for Ctrl (A1, A2: white histograms) and Ds Red2 L. am -hosting DLs (A3, A4) ( Ds Red2 + , black histograms) and amastigotes-free DLs ( Ds Red2 − , grey histograms). Mean fluorescence intensity was indicated for each histogram. Panel B: Statistical analysis of the BODIPY 493/503 MFI values. Mean BODIPY 493/503 fluorescence by MHC II + DLs was determined for n = 7 independent experiments. MFI in L. am -loaded cultures is shown for Ds Red2 + and Ds Red2 − DLs as black and grey bars, respectively. Statistical analyses were performed by the Mann Whitney test.

    Article Snippet: DL extracted RNA integrity control Total RNA was extracted from MHC II+ DLs (RNeasy+ Mini-Kit, Qiagen) and its quality and concentration was determined using a NanoDrop ND-1000 micro-spectrophotometer (Kisker, http://www.kisker-biotech.com ) and an Agilent-2100 Bioanalyzer (Agilent, http://www.chem.agilent.com ).

    Techniques: Fluorescence, Incubation, Staining, MANN-WHITNEY

    Flow cytometric analysis of CD36 expression in C57BL/6 DLs hosting live L. am amastigotes. L. am amastigotes were added to DL cultures at a ratio of 5 amastigotes per DL. Twenty-four hours later, DLs were detached and stained successively with anti- MHC II- PE-Cy5 and CD36-PE conjugated mAbs. As the fluorescence of transgenic Ds Red2 amastigotes was quashed by the fixation step, we used 2A3-26 mAb generated in our laboratory to image intact amastigotes inside their parasitophorous vacuoles. Thus, after fixation, the DLs were first permeabilized then labelled with alexafluor480-conjugated 2A3-26 mAbs before being analysed by FCM. This analysis was performed on gated MHC II + DLs. The central dot plot shows CD36 expression and the presence of intracellular parasites. The profiles of CD36 expression and mean CD36 fluorescence value are shown for 2A3-26 + DLs (red gate) and 2A3-26 − DLs (black gate).

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Reprogramming Neutral Lipid Metabolism in Mouse Dendritic Leucocytes Hosting Live Leishmania amazonensis Amastigotes

    doi: 10.1371/journal.pntd.0002276

    Figure Lengend Snippet: Flow cytometric analysis of CD36 expression in C57BL/6 DLs hosting live L. am amastigotes. L. am amastigotes were added to DL cultures at a ratio of 5 amastigotes per DL. Twenty-four hours later, DLs were detached and stained successively with anti- MHC II- PE-Cy5 and CD36-PE conjugated mAbs. As the fluorescence of transgenic Ds Red2 amastigotes was quashed by the fixation step, we used 2A3-26 mAb generated in our laboratory to image intact amastigotes inside their parasitophorous vacuoles. Thus, after fixation, the DLs were first permeabilized then labelled with alexafluor480-conjugated 2A3-26 mAbs before being analysed by FCM. This analysis was performed on gated MHC II + DLs. The central dot plot shows CD36 expression and the presence of intracellular parasites. The profiles of CD36 expression and mean CD36 fluorescence value are shown for 2A3-26 + DLs (red gate) and 2A3-26 − DLs (black gate).

    Article Snippet: DL extracted RNA integrity control Total RNA was extracted from MHC II+ DLs (RNeasy+ Mini-Kit, Qiagen) and its quality and concentration was determined using a NanoDrop ND-1000 micro-spectrophotometer (Kisker, http://www.kisker-biotech.com ) and an Agilent-2100 Bioanalyzer (Agilent, http://www.chem.agilent.com ).

    Techniques: Flow Cytometry, Expressing, Staining, Fluorescence, Transgenic Assay, Generated