GSM7566639: HUVEC-CVA10-0h; Homo sapiens; miRNA-Seq
Three replicates of CV-A10-infected cells at different time were mixed together and used for RNA extraction. In brief, total RNA isolation was conducted using a TRIzol™ reagent (Invitrogen, USA) according to manufacturer's guidelines. The concentration of total RNA was determined using a NanoDrop 2000 spectrophotometer. The RNA purity and the RNA integrity number (RIN) were assessed using an Agilent 2100 bioanalyzer. When the extracted RNA reached “An RIN score≥7.0 and rRNA 28S/18S≥1.6”, it was required in the following study. Total RNA was purified by polyacrylamide gel electrophoresis (PAGE) to enrich 15~35 nt molecules, then proprietary adapters were ligated to the 5' and 3' terminal of the RNA and the samples were used as templates for cDNA synthesis. The cDNA was amplified using the appropriate number of PCR cycles to produce sequencing libraries, which were subsequently subjected to the proprietary Solexa sequencing-by-synthesis method using the Illumina Genome Analyzer (SanDiego, USA). Sequencing was carried out at the ANOROAD Genome Inc. (Beijing, China).