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. 2014 Jun 24;16(8):871–879. doi: 10.1002/ejhf.116

Figure 1.

Figure 1

Schematic diagram illustrating the strategy for identification and validation of candidate microRNAs (miRs). For identification of candidate miRs we used a screening cohort of eight patients with paired plasma samples from before and after 6 months of left ventricular assist device therapy. Total RNA was extracted, and this was pooled into four test samples for each time point. We used a real-time quantitative polymerase chain reaction (RT-qPCR) microarray for unbiased expression profiling. Amplification failed for one sample pair and therefore this was excluded from analysis, giving a final n = 6 for the screening cohort. The microarray yielded 12 candidate miRs. These underwent validation using TaqMan RT-qPCR for greater specificity to minimize false-positive results, yielding miR-1202 and miR-483-3p as candidate miRs of interest. After fresh RNA extraction, these two miRs were quantified in the test cohort of all 73 samples (53 plasma and 20 LV myocardial tissue). All TaqMan reactions in the screening and test cohorts were performed in duplicate. See the Methods for more details.