GSM2691336: sh1; Mus musculus; RNA-Seq
Soleus muscles were removed, snap frozen in liquid nitrogen, and totalRNA was harvested using miRVana miRNA Isolation Kit (Ambion, Life Technologies). Illumina TruSeq RNA Sample Prep Kit was used with 1 ug of total RNA for the construction of sequencing libraries. After poly-A selection (using poly-T oligo-attached magnetic beads), mRNA was purified and fragmented using divalent cations under elevated temperature. The RNA fragments underwent reverse transcription using random primers. This is followed by second strand cDNA synthesis with DNA Polymerase I and RNase H. After end repair and A-tailing, indexing adapters were ligated. The products were then purified and, using 10 µl template, amplified (14 PCR cycles) to create the final cDNA libraries.