GSM5741307: WT1; Mus musculus; RNA-Seq
Total RNA was prepared using TRIzol Reagent (Invitrogen). DNase I was used to digest DNA in total RNA and rRNA was removed by hybridization of complementary DNA oligos to rRNA followed by degradation of the RNA:DNA hybrids using RNase H. Purified mRNA from previous steps was fragmented into small pieces and then used to prepare mRNA sequencing library. For quality control, libraries were validated using Agilent Technologies 2100 bio-analyzer. The final library was amplified via rolling circle amplification with phi29 DNA polymerase (Thermo Fisher Scientific, MA, USA), which could result in DNA nanoballs (DNB) that had more than 300 copies of one original molecule. DNBs were then loaded into the patterned nanoarray and pair end 150 bases reads were generated on a BGISEQ500 platform (BGI Shenzhen, China).