GSM8446947: PC3U LY 2157299; Homo sapiens; RNA-Seq
RNA was extracted from PC3U cells or A549 cells treated with or without TGFβ1 using the AllPrep DNA/RNA/Protein Mini Kit (QIAGEN, Cat. 80004) After the QC procedures, mRNA is enriched using oligo(dT) beads. The mRNA is then fragmented randomly in fragmentation buffer, followed by cDNA synthesis using random hexamersand 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.