Bulk RNA-sequencing of human neuroblastoma cell line SK-N-SH following cisplatin or JQ1 treatment
Bulk-RNA sequencing of human Neuroblastoma cell line SK-N-SH. Cells were treated with Cisplatin (48hrs treatment) or JQ1 (7 days treatment), following which cells were FACS sorted based on CD44 expression in high, intermediate, or low. The library construction method and sequencing process are carried out according to the following steps: oligo dT beads were used to enrich mRNA with poly A tail; mRNA molecules were fragmented and the fragmented mRNA was synthesized into first strand cDNA using random primers; the second strand cDNA was synthesized with dUTP instead of dTTP; the cDNA was subjected to end-repair and 3' adenylated; adaptors were ligated to the ends of the 3' adenylated cDNA fragments; the U-labelled second-strand template was digested with Uracil-DNA-Glycosylase and PCR amplification was performed; library underwent quality control and circularization; the library was amplified to make DNA nanoball and sequenced on DNBSEQ platform.