title: RNA-Seq of mhaESC
design: Total RNA was extracted by grinding tissue in TRIzol reagent (TIANGEN) on dry ice and processed following the protocol provided by the manufacturer.The poly-A RNAs were enriched from total RNA using Dynabeads mRNA Purification Kit (Cat#61006, Invitrogen) and fragmented into small pieces using fragmentation reagent in MGIEasy RNA Library Prep Kit V3.1 (Cat# 1000005276, MGI). The first strand cDNA was synthesized using random primes and reverse transcriptase, followed by second strand cDNA synthesis. The synthesized cDNA was end-repaired, A-tailing added and ligated to the sequencing adapters according to library construction protocol. The cDNA fragments were amplified by PCR and purified with MGIEasy DNA Clean beads (CAT#1000005279, MGI). Library was analyzed on the Agilent Technologies 2100 bioanalyzer. The double stranded PCR products were heat denatured and circularized by the splint oligo sequence in MGIEasy Circularization ModuleCAT#1000005260, MGI. The single strand circle DNA (ssCir DNA) were formatted as the final library. The qualified libraries were sequenced on DNBSEQ-T7RS platform.