title: GSM6281291: H1975 cells, sh-A, 3; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was harvested using an RNA Isolation Kit (ZYMO, Direct-zol RNA Miniprep Plus). Total RNA from each sample was treated with Oligo (dT) magnetic beads to select polyA mRNA. Target RNA fragments were reverse transcribed into double-strand cDNA (dscDNA) using N6 random primers. We then performed end repair on dscDNA fragments creating a phosphate at each 5′-end and an 'A' single nucleotide overhang at each 3′-end. Next, the adaptor with 'T' at 3′ end was ligated to the dscDNA, and ligation products were used for PCR amplification. The PCR products were denatured by heating, and single strand DNA was cyclized using a splint oligo and DNA ligase to format as the final library.