GSM7867753: Pax5_Wt_1_RNA; Mus musculus; RNA-Seq
RNA was extracted from frozen cell pellets using the Maxwell RSC simplyRNA Tissue Kit; For anti-Pax5 ChIP-Seq, sheared DNA was washed after the ChIP, crosslink was reversed and DNA was purified using NucleoSpin Gel and PCR Clean-up columns. For RNA-Seq, mRNA was enriched using oligo dT beads and fragmented. First strand cDNA was synthesized using random primers, and second strand cDNA was subjected to end-repair and 3' adenylated. Adaptors were ligated to the ends of the 3' adenylated cDNA fragments. U-labeled second-strand template was digested with UDG and amplified by PCR. After QC, library was circularized and amplified to make DNA nanoball before sequencing. For ChIP-Seq, IP-DNA and Input-DNA were 3' end repaired and adaptor-ligated before PCR amplification, purificaiton and sequencing.