GSM2973663: LV3; Mus musculus; RNA-Seq
Total RNA was harvested using Qiagen Rneasy mini kit following manufacturer's instructions. Poly-A containing mRNA molecules were purified using poly-T oligo attached magnetic beads and fragmented using divalent cations. The RNA fragments were transcribed into cDNA using  II Reverse Transcriptase (Invitrogen), followed by second strand cDNA synthesis using DNA Polymerase I and RNase H. Finally, cDNA fragments then have the addition of a single 'A' base and subsequent ligation of the adapter. The products were then purified and enriched by PCR using paired-end primers (Illumina) for 15 cycles to create the final cDNA library. RNA-seq libraries were prepared using the TruSeq Stranded mRNA Kit (Illumina) following the manufacturer's instructions.