GSM8121050: D2_5; Homo sapiens; RNA-Seq
RNA was isolated using miRNeasy Mini Kit (Qiagen, 217004) according to the manufacturer's guidelines polyA-tail enrichment was performed. Subsequently mRNA was randomly fragmented and cDNA synthesis proceeds through the use of random hexamers and the reverse transcriptase enzyme. After synthesis of first chain teh second chain was synthesized with addition of an Illumina buffer along with dNTPs, RNAseH and polymerase I from E. coli by Nick translation. resulting products went through purification, end-repair, A-tailing and adapter ligation. Fragments of appropriate size are enriched by PCR where P5 and P7 primers are introduced and dinal products purified.