Total RNA was extracted using Trizol with Ambion's protocol. mRNA was purified with oligo-dT beads, and used for cDNA synthesis with random primers after a fragmentation step with magnesium followed by heat-catalyzed hydrolysis. The cDNA 3' ends were adenylated, followed by adaptor ligation and a 15-cycle PCR to enrich DNA fragments. cDNA libraries were generated using a v2 Guide Quantification of cDNA libraries was performed by using Kapa Biosystems primer premix kit with Illumina-compatible DNA primers. The cDNA libraries were pooled at a final concentration 1.8 pM. Single-read sequencing was performed on Illumina Genome Analyzer IIx and NextSeq 500.
GEO Accession
GSM4696538