title: Whole genome sequencing of Equus kiang: adult male
design: Poly-A RNA was enriched from total RNA using the Dynabeads mRNA Purification Kit (Cat#61006, Invitrogen), and the mRNA was fragmented into small segments using the fragmentation reagent from the MGIEasy RNA Library Prep Kit V3.1 (Cat#1000005276, MGI). First-strand cDNA was synthesized using random primers and reverse transcriptase, followed by the synthesis of second-strand cDNA. According to the library construction protocol, the synthesized double-stranded cDNA (dscDNA) underwent end repair, A-tailing, and adapter ligation. Fragment size selection was performed using MGIEasy DNA Clean Beads (CAT#1000005279, MGI), and finally, PCR amplification was carried out to obtain the transcriptome library. The transcriptome library was quality-checked using an Agilent 2100 Bioanalyzer. Double-stranded DNA with adapter sequences was thermally denatured to form single-stranded DNA. Circularization primers from the MGIEasy Circularization Module (CAT#1000005260, MGI) were used to complementarily pair with both ends of the single-stranded DNA, forming single-stranded circular DNA (ssCir DNA). The concentration of the circularized product was detected before loading onto the sequencer, and qualified libraries were sequenced on the MGI-T7 sequencer.