RNAseq of experiment1
mRNA was purified from total RNA using poly-T oligo-attached magnetic beads.Fragmentation was carried out using divalent cations under elevated temperature. First strand strand was synthesized by random hexamers, 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 NGSR 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).