rneasy mini kits  (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
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    RNeasy Mini Kit
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    Structured Review

    Qiagen rneasy mini kits
    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 kits/product/Qiagen
    Average 99 stars, based on 744 article reviews
    Price from $9.99 to $1999.99
    rneasy mini kits - by Bioz Stars, 2020-05
    99/100 stars

    Images

    1) Product Images from "Tetraspanin CD63 is a regulator of HIV-1 replication"

    Article Title: Tetraspanin CD63 is a regulator of HIV-1 replication

    Journal: International Journal of Clinical and Experimental Pathology

    doi:

    CD63 mRNA down regulation by siRNA in human MDMs which are challenged with different concentration of HIV-1. A. MDMs (5 × 10 5 cells/well) were plated in 24-well plates and transfected with 50 nM siRNAs (CD63, CD4, and ERBB2IP). For controls, cells were treated with AZT (1 mM) or raltegravir (20 mM). Controls also included untreated cells or cells infected with HIV-1 SX (m.o.i. = 0.02) only. Total mRNA was isolated from each well using the Qiagen RNeasy kit 48 h after transfection. Quantitative RT-PCR was used to determine relative CD63 expression levels, normalizing to GAPDH expression as an internal control. B. MDMs were challenged with different concentration of HIV-1 SX (m.o.i. = 0.6, 0.2. 0.06 and 0.02, respectively). Supernatants were harvested for p24 detection on day 7 post-infection for MDMs using p24 Capture ELISA kit (ImmunoDiagnostics, Woburn, MA).
    Figure Legend Snippet: CD63 mRNA down regulation by siRNA in human MDMs which are challenged with different concentration of HIV-1. A. MDMs (5 × 10 5 cells/well) were plated in 24-well plates and transfected with 50 nM siRNAs (CD63, CD4, and ERBB2IP). For controls, cells were treated with AZT (1 mM) or raltegravir (20 mM). Controls also included untreated cells or cells infected with HIV-1 SX (m.o.i. = 0.02) only. Total mRNA was isolated from each well using the Qiagen RNeasy kit 48 h after transfection. Quantitative RT-PCR was used to determine relative CD63 expression levels, normalizing to GAPDH expression as an internal control. B. MDMs were challenged with different concentration of HIV-1 SX (m.o.i. = 0.6, 0.2. 0.06 and 0.02, respectively). Supernatants were harvested for p24 detection on day 7 post-infection for MDMs using p24 Capture ELISA kit (ImmunoDiagnostics, Woburn, MA).

    Techniques Used: Concentration Assay, Transfection, Infection, Isolation, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

    Effects of CD63 silencing on HIV-1 replication in human PBLs and DCs. A. PBLs (1 × 10 5 cells/well) were plated in triplicate in 24-well plates and transfected with 50 nM siRNAs (CD63, CD4, and ERBB2IP). Controls included untreated cells or cells treated with virus infection only. Total mRNA was isolated from each well using the Qiagen RNeasy kit 48 h after transfection. Quantitative RT-PCR was used to determine relative CD63 expression levels, normalizing to GAPDH expression as an internal control. B. PBLs cells (1 × 10 5 cells/well) or (C) DCs cells (5 × 10 5 cells/well) were plated in triplicate in 24-well plates and transfected with 50 nM siRNAs (CD63, CD4, and ERBB2IP). Controls included untreated cells or cells treated with virus infection only. Forty eight hours post-transfection, cells were infected with HIV-1 89.6 (m.o.i. = 0.02). Supernatants were harvested for p24 detection on day 5 post-infection for PBLs cells, and on day 7 post-infection for DCs using p24 Capture ELISA kit (ImmunoDiagnostics, Woburn, MA). C. Cell lysates from MDMs on day 7 post-infection were also harvested for detection of intracellular p24. *P
    Figure Legend Snippet: Effects of CD63 silencing on HIV-1 replication in human PBLs and DCs. A. PBLs (1 × 10 5 cells/well) were plated in triplicate in 24-well plates and transfected with 50 nM siRNAs (CD63, CD4, and ERBB2IP). Controls included untreated cells or cells treated with virus infection only. Total mRNA was isolated from each well using the Qiagen RNeasy kit 48 h after transfection. Quantitative RT-PCR was used to determine relative CD63 expression levels, normalizing to GAPDH expression as an internal control. B. PBLs cells (1 × 10 5 cells/well) or (C) DCs cells (5 × 10 5 cells/well) were plated in triplicate in 24-well plates and transfected with 50 nM siRNAs (CD63, CD4, and ERBB2IP). Controls included untreated cells or cells treated with virus infection only. Forty eight hours post-transfection, cells were infected with HIV-1 89.6 (m.o.i. = 0.02). Supernatants were harvested for p24 detection on day 5 post-infection for PBLs cells, and on day 7 post-infection for DCs using p24 Capture ELISA kit (ImmunoDiagnostics, Woburn, MA). C. Cell lysates from MDMs on day 7 post-infection were also harvested for detection of intracellular p24. *P

    Techniques Used: Transfection, Infection, Isolation, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

    2) Product Images from "Dysregulation of matrix metalloproteinases and their tissue inhibitors is related to abnormality of left ventricular geometry and function in streptozotocin-induced diabetic minipigs"

    Article Title: Dysregulation of matrix metalloproteinases and their tissue inhibitors is related to abnormality of left ventricular geometry and function in streptozotocin-induced diabetic minipigs

    Journal: International Journal of Experimental Pathology

    doi: 10.1111/j.1365-2613.2008.00579.x

    mRNA levels of MMPs and TIMPs in aortic intima and left ventricular (LV) myocardium by RT-PCR. Total RNA of aortic intima and LV myocardium was extracted using RNeasy Mini Kits and RT was done. Semi-quantitative PCR amplification was performed to amplify
    Figure Legend Snippet: mRNA levels of MMPs and TIMPs in aortic intima and left ventricular (LV) myocardium by RT-PCR. Total RNA of aortic intima and LV myocardium was extracted using RNeasy Mini Kits and RT was done. Semi-quantitative PCR amplification was performed to amplify

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Amplification

    3) Product Images from "Survival, gene and metabolite responses of Litoria verreauxii alpina frogs to fungal disease chytridiomycosis"

    Article Title: Survival, gene and metabolite responses of Litoria verreauxii alpina frogs to fungal disease chytridiomycosis

    Journal: Scientific Data

    doi: 10.1038/sdata.2018.33

    Schematic overview of the transcriptomic analyses performed in the study, and the corresponding quality control measures and data outputs. Three tissues were collected from each frog (n=61) for transcriptomic analysis, yielding a total of n=181 samples (2 missing data). All tissue samples underwent RNA extraction, however skin and liver samples had their total RNA extracted with 5-Prime PerfectPure kit and were subject to a DNase step, skin samples were subject to a Proteinase K step, and spleen samples had their RNA extracted with Qiagen RNeasy minikit. All RNA samples were subject to quality control steps, library preparation and sequencing, producing raw sequence data for each sample. Tissue-specific transcriptomes were assembled de novo with Trinity 34 and functionally annotated with BLAST2GO 36 (with inbuilt quality control measures, including minimum e-value and percentage similarity), and henceforth all sample data were aligned and annotated based on these.
    Figure Legend Snippet: Schematic overview of the transcriptomic analyses performed in the study, and the corresponding quality control measures and data outputs. Three tissues were collected from each frog (n=61) for transcriptomic analysis, yielding a total of n=181 samples (2 missing data). All tissue samples underwent RNA extraction, however skin and liver samples had their total RNA extracted with 5-Prime PerfectPure kit and were subject to a DNase step, skin samples were subject to a Proteinase K step, and spleen samples had their RNA extracted with Qiagen RNeasy minikit. All RNA samples were subject to quality control steps, library preparation and sequencing, producing raw sequence data for each sample. Tissue-specific transcriptomes were assembled de novo with Trinity 34 and functionally annotated with BLAST2GO 36 (with inbuilt quality control measures, including minimum e-value and percentage similarity), and henceforth all sample data were aligned and annotated based on these.

    Techniques Used: RNA Extraction, Sequencing

    Related Articles

    RNA Extraction:

    Article Title: NF-κB promotes the stem-like properties of leukemia cells by activation of LIN28B
    Article Snippet: .. RNA extraction and qRT-PCR RNeasy Mini Kit (Qiagen Hilden, Germany) was purchased for RNA extraction. .. In brief, cDNA was synthesized by using the i-Script™ Reverse Transcription Supermix (Biorad, Hercules, CA, United States) from 1 µg of total RNA.

    Article Title: RNA extraction from self-assembling peptide hydrogels to allow qPCR analysis of encapsulated cells
    Article Snippet: .. RNA extraction from peptide hydrogels Attempts were made to extract RNA from cells using two commercial kits: one solution-based, TRI Reagent® (Sigma-Aldrich), and one column-based, 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).

    Isolation:

    Article Title: A preliminary analysis of hepatitis C virus in pancreatic islet cells
    Article Snippet: .. mRNA expression of HCV entry factors Total RNA from 3 different human islets donors was isolated using TRIzol reagent (Thermo Scientific) in combination with the RNeasy Mini kit (Qiagen) followed by DNase treatment. .. Five hundred nanograms of total RNA were retrotranscribed using the Superscript III kit (Thermo Scientific), and the cDNAs obtained were utilized as templates for quantitative real-time RT-PCR analysis of CD81 (72 base pair [bp]), occludin (63 bp), claudin-1 (101 bp), and SR-B1 (50 bp), as well as the housekeeping gene GAPDH (94 bp). cDNA was run on an AbiPRISM 7300 fast real-time cycler using the power SYBR Green real-time PCR master mix kit and quantified by built-in SYBR Green Analysis.

    Article Title: PTK787/ZK22258 attenuates stellate cell activation and hepatic fibrosis in vivo by inhibiting VEGF signaling
    Article Snippet: .. Total RNA was isolated from liver tissues and HSCs using RNeasy Mini kit (Qiagen Inc., Hilden, Germany), according to the manufacturer’s instruction. .. For cDNA synthesis, Taqman reverse transcription reagents were used as described in the manufacture’s protocol (PE Applied Biosystems, Foster City, CA, USA).

    Quantitative RT-PCR:

    Article Title: NF-κB promotes the stem-like properties of leukemia cells by activation of LIN28B
    Article Snippet: .. RNA extraction and qRT-PCR RNeasy Mini Kit (Qiagen Hilden, Germany) was purchased for RNA extraction. .. In brief, cDNA was synthesized by using the i-Script™ Reverse Transcription Supermix (Biorad, Hercules, CA, United States) from 1 µg of total RNA.

    Article Title: PDCD6 is an independent predictor of progression free survival in epithelial ovarian cancer
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) RNeasy Mini Kits (QIAGEN Inc., Valencia, CA) were used to extract total RNA from cells and tissue. .. RNA (5 ng) was reverse transcribed to cDNA using a SuperScript First-Strand Synthesis System for RT-PCR (Invitrogen Corp., Carlsbad, CA).

    Purification:

    Article Title: Quality Control of RNA Preservation and Extraction from Paraffin-Embedded Tissue: Implications for RT-PCR and Microarray Analysis
    Article Snippet: .. RNA Purification and Complementary DNA (cDNA) Synthesis Where indicated, the total RNA obtained by Trizol extraction was purified by processing with RNeasy Mini Kit (Qiagen) or RNeasy miRNA Mini Kit (Qiagen) according to manufactureŕs instructions. .. After extraction and purification, the RNA yield and purity was determined by measuring absorbance at 260 nm/280 nm on a Nanodrop spectrophotometer (Thermo Fisher Scientific).

    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: PDCD6 is an independent predictor of progression free survival in epithelial ovarian cancer
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) RNeasy Mini Kits (QIAGEN Inc., Valencia, CA) were used to extract total RNA from cells and tissue. .. RNA (5 ng) was reverse transcribed to cDNA using a SuperScript First-Strand Synthesis System for RT-PCR (Invitrogen Corp., Carlsbad, CA).

    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 ). ..

    Expressing:

    Article Title: A preliminary analysis of hepatitis C virus in pancreatic islet cells
    Article Snippet: .. mRNA expression of HCV entry factors Total RNA from 3 different human islets donors was isolated using TRIzol reagent (Thermo Scientific) in combination with the RNeasy Mini kit (Qiagen) followed by DNase treatment. .. Five hundred nanograms of total RNA were retrotranscribed using the Superscript III kit (Thermo Scientific), and the cDNAs obtained were utilized as templates for quantitative real-time RT-PCR analysis of CD81 (72 base pair [bp]), occludin (63 bp), claudin-1 (101 bp), and SR-B1 (50 bp), as well as the housekeeping gene GAPDH (94 bp). cDNA was run on an AbiPRISM 7300 fast real-time cycler using the power SYBR Green real-time PCR master mix kit and quantified by built-in SYBR Green Analysis.

    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).

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  • 99
    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 8429 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy mini kit/product/Qiagen
    Average 99 stars, based on 8429 article reviews
    Price from $9.99 to $1999.99
    rneasy mini kit - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

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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