GSM3954942
GSM3954942: S31_171_KO; Mus musculus; RNA-Seq
6mo Mss51 WT and KO animals were maintained under a 12:12 hour light:dark schedule with ad libitum access to food and water. Quadriceps muscle were isolated from each mouse and frozen in cold isopentane (#M32631, Sigma-Aldrich). Muscle tissues were homogenized in TRIzol (15596018, Life Technologies) and 2 ug of total RNA was isolated using the Direct-zol RNA MiniPrep Kit (R2052, Zymo Research, Irvine, CA, USA) for the construction of sequncing libraries. After the QC procedures, mRNA from eukaryotic organisms was enriched using oligo(dT) beads. rRNA was removed using the Ribo-Zero kit that leaves the mRNA. First, the mRNA was fragmented randomly by adding fragmentation buffer, then the cDNA was synthesized by using mRNA template and random hexamers primer, after which a custom second-strand synthesis buffer (Illumina), dNTPs, RNase H and DNA polymerase I were added to initiate the second-strand synthesis. Second, after a series of terminal repairs, ligation and sequences adapter ligation, the double-stranded cDNA library was completed through size selection and PCR enrichment. Captured library was sequenced on Illumina HiSeq 4000 (Illumina) platform according to manufacturer's instructions for paired-end 150 bp sequencing with sequence coverage of 50 million read-pairs.
GEO
GSM3954942
GEO Accession
GSM3954942