2,6-DCP treatment
Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was quantified with Qubit and assessed with real-time PCR using a bioanalyzer for size distribution detection. Quantified libraries were pooled and sequenced on an Illumina platform according to the manufacturer's specifications.