ctrl_97hpf#3
The RNA sample was sent to Biomarker Technologies (http://www.biomarker.com.cn) for library construction and RNA sequencing. Briefly, mRNA molecules were enriched using Oligo (dT) magnetic beads and randomly fragmented. First strand cDNA synthesis was performed using random hexamers. Second strand cDNAs were synthesized by adding dNTPs, RNase and DNA polymerase. The double stranded cDNAs were purified using AMPure XP beads. After end filling, A-tailing and sequencing adaptor ligation, cDNA fragments of desired size were selected using AMPure XP beads. Finally, the sequencing library was generated by PCR amplification. Library quantification was performed using a Qubit 2.0 and the insert size of library was analyzed by an Agilent 2102 Bioanalyzer system.