title: GSM2064258: 1st 12h JQ1(+) 0.2 µM; Mus musculus; RNA-Seq
library_construction_protocol: RNA was isolated using  RNeasy Mini Kit (Qiagen, Valencia, CA). The samples were quantified and more than 2 μg of total RNA for each sample was sent to BGI for library construction. i. DNase I digest: DNase I degrade double-stranded and single-stranded DNA in RNA samples.  ii. mRNA isolation: Poly (A)-containing mRNA molecules were purified from total RNA using poly (T) oligo-attached magnetic beads.  iii. mRNA fragment: mRNA were fragmented into small pieces using fragmentation reagent.  iv. cDNA synthesis: First-strand cDNA was generated using random hexamer-primed reverse transcription. This was followed by a second-strand cDNA synthesis and a cDNA purify using purification Kit.  v. End repair, add A and adaptor ligation: The synthesized cDNA was subjected to end-repair and then was 3' adenylated. Adaptors were ligated to the ends of these 3' adenylated cDNA fragments. Ligation products were purified with the magnetic beads, and dissolved in the appropriate amount of Epstein-Barr solution.  vi. RCR: This process was to amplify those cDNA fragments with adaptors from previous step. PCR products were purified with the magnetic beads, and dissolved in the appropriate amount of Epstein-Barr solution.  vii. Library quality control: Library was validating on the Agilent Technologies 210 RNA-Seq. Single-end 50bp (SE50) sequencing was done on the HiSeq2000 sequencing platform (Illumina, Inc, CA, USA) according to the manufacturer's protocol.
attributes.tag: GEO Accession
attributes.value: GSM2064258