Journal: PLoS ONE
Article Title: Stability of Circulating Blood-Based MicroRNAs – Pre-Analytic Methodological Considerations
Figure Lengend Snippet: Different blood tubes for miRNA analysis. miRNA was isolated and levels of miR-1, miR-21 and miR-29b from either; EDTA-plasma, citrate-plasma, lithium-heparin-plasma, or serum fraction, were detected by RT-qPCR. Values were normalized to a spike-in control, cel-miR-39. Normalized average ΔC T values for each are shown, demonstrating amplification from all sources, except lithium-heparin-plasma. In the remaining samples, the levels of miR-21 and miR-29b did not vary significantly between the blood tubes. Li = lithium, ND = not detectable. A) Copenhagen Cohort. B) Munich Cohort.
Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) qRT-PCR was perfomed using TaqMan Gene Expression MasterMix (Life Technologies, USA) and TaqMan probes (Life Technologies, USA) for hsa-miR-1 (Assay-ID: 000385), hsa-miR-21 (Assay-ID:000397), hsa-miR-29b (Assay-ID: 000413) and cel-miR-39 (Assay-ID: 000200) according to manufacturer’s instructions under sterile conditions using either an Applied Biosystems 7300 Real-Time PCR System (Copenhagen samples) or a CFX 96 C1000 touch Realtime Cycler (Bio Rad, Germany) (Munich samples).
Techniques: Isolation, Quantitative RT-PCR, Amplification