Extraction and Reverse Transcription of miRNA from Coronary Atherosclerotic Plaques

Total RNA was extracted from 23 tissue sections obtained from 12 coronary atherosclerotic plaques (Table 1) using the Roti-Quick-Kit (Carl Roth GmbH, Karlsruhe, Germany) following the manufacturer’s instructions and as described previously^{10 (link),12 (link)}. RNA concentrations were measured using an Eppendorf BioPhotometer. In addition, we used the SABiosciences RT² qPCR-Grade miRNA Isolation Kit (SABiosciences Corporation, Frederick, MD, USA) to enrich miRNA from 40 µg of total RNA of each sample according to manufacturer’s instructions. This kit combines a phenol/chloroform-based extraction method with a silica membrane spin column technology. MiRNA was then reverse transcribed using the SABiosciences RT² miRNA FirstStrand Kit according to the manufacturer’s protocol and as described previously^{10 (link),12 (link)}.