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
    Price:
    314
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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 160 article reviews
    Price from $9.99 to $1999.99
    rneasy mini kits - by Bioz Stars, 2020-09
    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: 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: 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). ..

    Synthesized:

    Article Title: Amino Acids as Signaling Molecules Modulating Bone Turnover
    Article Snippet: .. 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). .. For PCR reactions the “Platinum PCR SuperMix” (Invitrogen) was used (94°C, 30 sec; 57° C, 30 sec; 72° C, 1 min for 35 cycles).

    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: 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).

    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). ..

    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) SynthesisWhere 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: 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).

    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.

    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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  • 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 36596 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 36596 article reviews
    Price from $9.99 to $1999.99
    rneasy mini kit - by Bioz Stars, 2020-09
    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

    Quantitative RT-PCR analysis of PAI-1 mRNA levels in patients. The total RNA was harvested from the peripheral blood mononuclear cells using an RNeasy kit according to the manufacturer’s instructions. The RT-PCR experiments were repeated at least 3 times. RNA was reverse transcribed into cDNA using random primers in a Reverse Transcription II system according to the manufacturer’s instructions. The expression of PAI-1 mRNA was quantified by quantitative PCR using an ABI Prism Sequence Detection System. Template-negative and RT-negative conditions were used as controls. Amplification of the endogenous GAPDH cDNA was monitored. The levels (mean value) of PAI-1 transcripts in patients were calculated. RT-PCR, reverse-transcription-PCR, PAI-1, plasminogen activator inhibitor-1.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Therapeutic effects of calcium dobesilate on diabetic nephropathy mediated through reduction of expression of PAI-1

    doi: 10.3892/etm.2012.755

    Figure Lengend Snippet: Quantitative RT-PCR analysis of PAI-1 mRNA levels in patients. The total RNA was harvested from the peripheral blood mononuclear cells using an RNeasy kit according to the manufacturer’s instructions. The RT-PCR experiments were repeated at least 3 times. RNA was reverse transcribed into cDNA using random primers in a Reverse Transcription II system according to the manufacturer’s instructions. The expression of PAI-1 mRNA was quantified by quantitative PCR using an ABI Prism Sequence Detection System. Template-negative and RT-negative conditions were used as controls. Amplification of the endogenous GAPDH cDNA was monitored. The levels (mean value) of PAI-1 transcripts in patients were calculated. RT-PCR, reverse-transcription-PCR, PAI-1, plasminogen activator inhibitor-1.

    Article Snippet: The total RNA was harvested from peripheral blood mononuclear cells using the RNeasy kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions.

    Techniques: Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Expressing, Real-time Polymerase Chain Reaction, Sequencing, Amplification, Polymerase Chain Reaction

    2′- O methylation of internal adenosine. (A) Incorporation of 3 H-methyl into polyA. Homopolymer RNAs (1 µg) were incubated with 2 µg of DENV-4 MTase in the presence of [ 3 H-methyl]-SAM. After the methylation reaction, the un-incorporated [ 3 H-methyl]-SAM was removed by RNeasy kit. The amount of 3 H-methyl incorporation was measured by a MicroBeta counting. (B) SPA-based methylation analysis of oligo (A) 12 , (Am) 12 , and (m 6 ,m 6 A) 12 . All three RNA oligos were 3′-end biotinylated to facilitate SPA analysis. Am indicates that the 2′-OH of adenosine is methylated. m 6 ,m 6 A indicates that the amino N 6 position of adenosine is double methylated. (C) SPA-based methylation analysis of DENV-1 RNA. pppA-RNAs, representing the 5′ 211 nt of DENV-1 genome, were in vitro transcribed using biotinylated-CTP plus unmodified ATP, 2′- O -methyladenosine triphosphate (AmTP), or N 6 methyl adenosine triphosphate (m 6 ATP). The transcription reactions generated pppA-RNA, ppp(Am)-RNA, and ppp(m 6 A)RNA, respectively. The RNAs were then subjected to SPA-based internal methylation analysis. Average results and standard deviations from three independent experiments are presented.

    Journal: PLoS Pathogens

    Article Title: 2?-O Methylation of Internal Adenosine by Flavivirus NS5 Methyltransferase

    doi: 10.1371/journal.ppat.1002642

    Figure Lengend Snippet: 2′- O methylation of internal adenosine. (A) Incorporation of 3 H-methyl into polyA. Homopolymer RNAs (1 µg) were incubated with 2 µg of DENV-4 MTase in the presence of [ 3 H-methyl]-SAM. After the methylation reaction, the un-incorporated [ 3 H-methyl]-SAM was removed by RNeasy kit. The amount of 3 H-methyl incorporation was measured by a MicroBeta counting. (B) SPA-based methylation analysis of oligo (A) 12 , (Am) 12 , and (m 6 ,m 6 A) 12 . All three RNA oligos were 3′-end biotinylated to facilitate SPA analysis. Am indicates that the 2′-OH of adenosine is methylated. m 6 ,m 6 A indicates that the amino N 6 position of adenosine is double methylated. (C) SPA-based methylation analysis of DENV-1 RNA. pppA-RNAs, representing the 5′ 211 nt of DENV-1 genome, were in vitro transcribed using biotinylated-CTP plus unmodified ATP, 2′- O -methyladenosine triphosphate (AmTP), or N 6 methyl adenosine triphosphate (m 6 ATP). The transcription reactions generated pppA-RNA, ppp(Am)-RNA, and ppp(m 6 A)RNA, respectively. The RNAs were then subjected to SPA-based internal methylation analysis. Average results and standard deviations from three independent experiments are presented.

    Article Snippet: For each time point, total cellular RNA was extracted using RNeasy kit (Qiagen).

    Techniques: Methylation, Incubation, In Vitro, Generated