GSM3928327: Brg1WT2; Mus musculus; RNA-Seq
Total RNA was extracted using Trizol reagent (Invitrogen).Total RNA sample is digested by DNaseІ (NEB), and purified by oligo-dT beads (Dynabeads   mRNA purification kit, Invitrogen) Construction of the cDNA library and transcriptome sequencing were performed by BGI Genomics, BGI-Shenzhen, as following. First-strand cDNA was generated by SuperScript III reverse transcriptase (Thermo Fisher Scientific). Free primers were removed using Exonuclease I (New England Biolabs). 3' poly (A) tailing was done by adding terminal deoxynucleotidyl transferase (Thermo Fisher Scientific). Then, the second-strand cDNA was synthesized and the cDNA was amplified by PCR. PCR products were purified with Ampure XP Beads (Beckman Coulter). After that, second round of PCR was performed and PCR products were fragmented by a sonicator (Covaris). End repaired was performed and A-tailing was added. Adaptor ligation was done and the cDNA was further amplified by PCR and purified with Ampure XP Beads. Fragment sizes were checked using Agilent 2100 bioanalyzer instrument (Agilent DNA 1000 Reagents) and the library was quantified by real-time quantitative PCR (QPCR) (TaqMan Probe). The qualified libraries were amplified on cBot to generate the cluster on the flowcell (TruSeq PE Cluster Kit V3–cBot–HS，Illumina). And the amplified flowcell was sequenced single end on the HiSeq 2000 System (TruSeq SBS KIT-HS V3，Illumina) with reading length of 90bp.
GEO
GSM3928327
GEO Accession
GSM3928327