RNA_seq,MK,rep1
Sequencing libraries were generated according to the following steps. Firstly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. Fragmentation was carried out using divalent cations under elevated temperature in fragmentation buffer. First strand was synthesized by randomhexamers, and the second cDNA strand was synthesized by adding buffer, dNTPs, RNase H and DNA polymerase I. Double stranded cDNA was repaired and A was added to the 3 'end. Hieff NGS DNA Selection Beads were used for purification and fragment Selection. After purification and fragment selection, the products were amplified and enriched by PCR. The target region library of the duplex was denatured, cycled, and digested to yield asingle-stranded circular DNA. Single-stranded circular DNA is amplified by arolling circle amplification (RCA), known as DNA nano balls (DNB). After library construction, Qubit was used for quantitative.