Journal: BMC Genomics

Article Title: Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells

doi: 10.1186/1471-2164-15-395

Figure Lengend Snippet: Assessment of the TLDA reproducibility. The TLDA reproducibility was assessed by testing, from one RNA sample, two separate qPCR amplifications from A - the same RT and pre-amplification product (RT1-d1 vs RT1-d2: Bland-Altman) and B - separate RT and pre-amplification reactions (RT1 (d1) vs RT2: Bland-Altman). RT with 150 ng of RNA isolated from 3×10 6 cells with Macherey-Nagel kit. Only miRNAs with Ct < 32 were considered.

Article Snippet: In that context, we performed an independent cross-comparison of three commercially available RNA extraction kits for miRNAs isolation from human blood samples (Qiagen and Norgen kits as well as the new NucleoSpin miRNAs Plasma kit from Macherey-Nagel). miRNAs were further profiled using the Taqman Low Density Array technology.

Techniques: Amplification, Isolation

Journal: BMC Genomics

Article Title: Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells

doi: 10.1186/1471-2164-15-395

Figure Lengend Snippet: Impact of the RNA quantity on miRNA TLDA profiles. miRNA TLDA profiles were compared using three different RNA quantities for RT (10, 100 and 300 ng), of the same RNA sample extracted from 3×10 6 PBMCs by Macherey-Nagel. A - Bland-Altman analysis: condition 100 ng of RNA for RT versus 10 ng. B - Bland-Altman analysis: condition 300 ng of RNA for RT versus 100 ng. C - Venn diagram with the detectable miRNAs. Only miRNAs with Ct < 32 were considered. The Pearson correlation coefficient is indicated.

Article Snippet: In that context, we performed an independent cross-comparison of three commercially available RNA extraction kits for miRNAs isolation from human blood samples (Qiagen and Norgen kits as well as the new NucleoSpin miRNAs Plasma kit from Macherey-Nagel). miRNAs were further profiled using the Taqman Low Density Array technology.

Techniques:

Journal: BMC Genomics

Article Title: Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells

doi: 10.1186/1471-2164-15-395

Figure Lengend Snippet: Comparison of miRNAs expression profiles from human PBMCs and serum. Human PBMCs (1×10 6 cells) and serum (300 μL) samples were extracted by Macherey-Nagel (MN) and Qiagen (Q) kits. A - Number of miRNAs detected from PBMCs RNA samples using TLDA cards. B - Correlation analysis from PBMCs samples. C - Bland-Altman analysis MN versus Q from PBMCs. D - Number of miRNAs detected from serum RNA samples using TLDA cards. E - Correlation analysis from serum samples. F - Bland-Altman analysis MN versus Q from serum. TLDA data were obtained from biological triplicate for each extraction kits. Analysis using mean Ct values of triplicate. Only miRNAs with Ct < 32 were considered for PBMCs. No Ct cut-off was applied for serum miRNAs.

Article Snippet: In that context, we performed an independent cross-comparison of three commercially available RNA extraction kits for miRNAs isolation from human blood samples (Qiagen and Norgen kits as well as the new NucleoSpin miRNAs Plasma kit from Macherey-Nagel). miRNAs were further profiled using the Taqman Low Density Array technology.

Techniques: Comparison, Expressing, Extraction

Journal: BMC Genomics

Article Title: Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells

doi: 10.1186/1471-2164-15-395

Figure Lengend Snippet: Assessment of bias in RNA isolation using Macherey-Nagel kit from 300 or 600 μL serum. A - Bland-Altman analysis 600 versus 300 μL of serum. B - Plot of the difference in Ct values of the two conditions (x-axis) and the GC content of the miRNAs detected in these two settings (y-axis). The Pearson correlation coefficient is indicated. The GC content corresponds to the percentage of GC in the sequence of mature miRNAs. TLDA datas from biological duplicate. Analysis using mean CT values of common miRNAs, without Ct cut-off.

Article Snippet: In that context, we performed an independent cross-comparison of three commercially available RNA extraction kits for miRNAs isolation from human blood samples (Qiagen and Norgen kits as well as the new NucleoSpin miRNAs Plasma kit from Macherey-Nagel). miRNAs were further profiled using the Taqman Low Density Array technology.

Techniques: Isolation, Sequencing

Journal: BMC Genomics

Article Title: Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells

doi: 10.1186/1471-2164-15-395

Figure Lengend Snippet: Assessment of bias in RNA isolation from PBMCs. Assessment of bias in RNA isolation from PBMCs using the Macherey-Nagel (MN) kit, comparison of extraction from 3×10 6 and 1×10 6 cells with a same amount of RNA for RT (130 ng). A - Bland-Altman analysis 3×10 6 versus 1x10 6 cells. B - Plot of the difference in Ct values of the two conditions (x-axis) and the GC content of miRNAs detected in these two settings (y-axis). C - Plot of the difference in Ct values of the conditions 300 vs 100 ng (x-axis) and the GC content of the miRNAs detected in these two settings (y-axis). The Pearson correlation coefficient is indicated. TLDA datas from biological duplicate. Analysis using mean CT values of common miRNAs. Only miRNAs with Ct < 32 were considered.

Article Snippet: In that context, we performed an independent cross-comparison of three commercially available RNA extraction kits for miRNAs isolation from human blood samples (Qiagen and Norgen kits as well as the new NucleoSpin miRNAs Plasma kit from Macherey-Nagel). miRNAs were further profiled using the Taqman Low Density Array technology.

Techniques: Isolation, Comparison, Extraction