title: no applicate
design: <1 g RNA from the sample was used to prepare cDNA libraries using the NEBNext Ultra RNA Library Prep Kit for Illumina. rRNA was removed using the Ribo-Zero kit. mRNA was purified from total RNA using poly-T oligo-bound magnetic beads and randomly fragmented by addition of fragmentation buffer. cDNA was synthesized using an mRNA template and a random hexamer primer. Then, a custom buffer for second-strand synthesis (Illumina), dNTPs, RNase H, and DNA polymerase I were added to start second-strand synthesis. After a series of terminal repair, ligation, and sequencing adapter ligation reactions, the double-stranded cDNA library was completed by size selection and PCR enrichment