GSM3581364: 48h_2mg_CKI3_Rep3_Batch1; Homo sapiens; ssRNA-seq
Total RNA was isolated by using an RNA extraction kit (Thermo Fisher Scientific) according to the manufacturer's instructions and RNA samples were quantified and quality determined using a Bioanalyzer at the Cancer Genome Facility of the Australian Cancer Research Foundatin (Australia). RNA samples with RNA integrity number (RINs) > 7.0 were sent to be sequenced at Novogene (China). Briefly, after QC procedures were performed, mRNA was isolated using oligo(dT) beads and randomly fragmented by adding fragmentation buffer, followed by cDNA synthesis primed with random hexamers. Next, a custom second-strand synthesis buffer (Illumina) , dNTPs, RNase H and DNA polymerase I were added for second-strand synthesis. After end repair, barcode ligation and sequencing adaptor ligation, the double-stranded cDNA library was size selected and PCR amplified. Sequencing was carried out on an Illumina HiSeq platform PE150 with paired-end 150 bp reads.