GSM8829525: Kelly cells, DpC, repeat 1; Homo sapiens; RNA-Seq
Treated cells were collected in 1ml of TRI reagent (Sigma-Aldrich) and the RNA was extracted as per the manufacturer's instructions The library preparation kit, NEBNext Ultra II RNA Library Prep Kit for Illumina(Cat No. 7770), was used to purify messenger RNA from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers. Then the second strand cDNA was synthesized using dTTP. The library was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification.  The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries were pooled and sequenced on Illumina platform NovaSeq 6000 with sequencing strategy PE150 and generated at least 50 Mb of paired-end raw reads per sample.