GSM5243509: DMSO; Homo sapiens; RNA-Seq
Total RNA was extracted using TRIzol Reagent (Invitrogen-Life Technologies, Waltham, Massachusetts, USA) following the manufacturer's instructions. The following sequencing and data analysis work using these RNA samples were performed by BGI-Shenzhen (Shenzhen, Guangdong, China). Briefly, total RNA was qualified and quantified using a NanoDrop and Agilent 2100 bioanalyzer (Thermo Fisher Scientific, Waltham, MA, USA). Oligo(dT)-attached magnetic beads were used to purify mRNA. Purified mRNA was fragmented into small pieces with fragment buffer at the appropriate temperature. Then, first-strand cDNA was generated using random hexamer-primed reverse transcription, followed by second-strand cDNA synthesis. Afterwards, A-Tailing Mix and RNA Index Adapters were added via incubation for end repair. The cDNA fragments obtained from the previous step were amplified by PCR, and the products were purified by Ampure XP Beads (BECKMAN, Brea, CA, USA) and then dissolved in EB solution. The product was validated on an Agilent Technologies 2100 bioanalyzer (Agilent, Santa Clara, CA, USA) for quality control. The double-stranded PCR products from the previous step were heated, denatured, and circularized by the splint oligo sequence to obtain the final library. Single-stranded circular DNA (ssCir DNA) was formatted as the final library.
GEO Accession
GSM5243509