GSM8527678: BMDM cells, M, WT1; Mus musculus; RNA-Seq
Total RNA was isolated using RNAeasy PLUS mini kit (QIAGEN) according to the manufacturer's instructions. Total RNA was submitted to Haplox international for library construction and RNA sequencing. mRNA molecules were purified from total RNA using oligo(dT)-attached magnetic beads. mRNA molecules were fragmented into small pieces using a fragmentation reagent. First-strand cDNA was generated using random hexamer-primed reverse transcription, followed by a second-strand cDNA synthesis. The synthesized cDNA was subjected to end-repair and then was 3' adenylated. Adapters were ligated to the ends of these 3' adenylated cDNA fragments. cDNA fragments were amplified with adapters from the previous step. PCR products were purified with AMPure XP Beads (AGENCOURT) and dissolved in EB solution. The library was validated on the Agilent Agilent 4200 Tapestation.