VY1 (CRX1511832; CRA022807)
Total RNA was extracted using Trizol according to the manufacturers instructions. The quality and quantity of the RNA were assessed using the Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit.From 5 ug of total RNA, mRNA was isolated using Dynabeads Oligo-dT through 2 rounds of purification. The purified mRNA was fragmented and then reverse-transcribed into cDNA using SuperScript II Reverse Transcriptase. The first-strand cDNA was then used to synthesize U-labeled DNA.The U-labeled Heat-labile UDG enzyme was used to remove the U-labeled regions. The resulting fragments were amplified by PCR.