GSM4007543: BAT in Young mouse, cold room (4°c) smp2; Mus musculus; RNA-Seq
Total RNA was isolated using TRIzol from the brown adipose tissue of C57BL6J mice in sterile condition. Total RNA integrity was checked using 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA) with an RNA integrity number (RIN) value greater than 7. For RNA-Seq, RNA libraries were prepared using the TruSeq RNA library preparation kit (Illumina, San Diego, CA, USA) with a modified protocol. Illumina TruSeq Stranded Total RNA Library Prep Kit was used for library prepartion. rRNA was removed by Ribo-Zero, a reagent consisting of biotinylated oligonucleotides that are complimentary to rRNA. The purified mRNA was cleaved and reverse-transcribed into first-strand cDNA using reverse transcriptase and random primers. The RNA template was removed, followed by second-strand cDNA synthesis. End repair, 3' adenylation and adapter ligation were performed. The purified cDNA templates were then enriched using PCR. The constructed libraries were 50 bp paired-end sequenced using an Illumina HiSeq 2000 sequencer.