GSM7876841: Caki-1 shControl_2; Homo sapiens; RNA-Seq
Cells were then collected into Trizol reagent (Invitrogen #15596026) in RNAase-free microcentrifuge tubes for subsequent RNA extraction. Total RNA was extracted as recommended by the manufacturer. Sample processing and sequencing were performed by BIG.inc (Shenzhen China). Briefly, messenger RNA was purified with magnetic beads conjugated with Oligo-dT and then fragmented with fragmentation buffer. Then first-strand cDNA was synthesized with random primer. After second-strand cDNA synthesis, A-tailing and RNA index adapters were added. The cDNA fragments were amplified by PCR and products were purified by Ampure XP Beads. The product was analyzed on the Agilent Technologies 2100 bioanalyzer for quality control. The PCR product was heat-denatured and circularized by the splint oligo sequence to get the final library. The final library was amplified with Phi29 DNA Polymerase to make DNA nanoball (DNB) which had more than 300 copies of one molecular. Pair-end 150 bases reads were generated on MGISEQ-2000RS platform (BGI, Shenzhen, China).