GSM9245954: Non-edited, CCAOBAP1-WT clone 9, rep3; Homo sapiens; RNA-Seq
CCAOs grown in BME domes were washed three times with PBS, lysed with QIAzol Lysing Reagent, and processed using the miRNeasy Mini Kit (Qiagen) according to the manufacturer's instructions Library construction was performed at the Novogene sequencing facilities, Cambridge, UK.  In short, purified mRNA was fragmented into small pieces with fragment buffer at appropriate temperature. Then First-strand cDNA was generated using random hexamer-primed reverse transcription, followed by a second-strand cDNA synthesis. afterwards, A-Tailing Mix and RNA Index Adapters were added by incubating to end repair. The cDNA fragments obtained from previous step were amplified by PCR, and products were purified by Ampure XP Beads, then dissolved in EB solution.The double stranded PCR products from previous step were heated denatured and circularized by the splint oligo sequence to get the final library. The single strand circle DNA (ssCir DNA) was formatted as the final library.