title: GSM2722290: RT-7; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was isolated from cell lines by Trizol(invitrogen).mRNA was enriched from total RNA with size selection using Megaclear kit (invitrogen) followed by two rounds of polyA selection using Dynabeads Oligo(dT)25 (invitrogen); RNA fraction under 200nt was collected from the flow through of Megaclear kit. RNA samples were dephosphorylated with PNK (NEB) and ligated to 3’ RNA linker using T4 RNA ligase2, truncated KQ (NEB). Reverse transcription was carried using different reverse transcriptase for different samples.  5’ adaptor was ligated to cDNA and then amplified by PCR. PCR products were purified by 8% TBE gel