GSM7061664: PEO1 cells, si1A1_3; Homo sapiens; RNA-Seq
RNA was isolated using Norgen Biotek RNA Clean-Up and Concentration Kit After the QC procedures, mRNA is enriched using oligo(dT) beads, then fragmented randomly in fragmentation buffer, followed by cDNA synthesis using random hexamers and reverse transcriptase. After first-strand synthesis, a custom second-strand synthesis buffer (Illumina) is added with dNTPs, RNase H and Escherichia coli polymerase I to generate the second strand by nick-translation. The final cDNA library is ready after a round of purification, terminal repair, A-tailing, ligation of sequencing adapters, size selection and PCR enrichment.