GSM5226192: Ucp2-wildtype genipin treatment replicate 3; Mus musculus; RNA-Seq
The TruSeq Stranded Total RNA library preparation kit from Illumina was used. This protocol involves the removal of ribosomal RNA (rRNA) using biotinylated, target-specific oligos combined with Ribo-Zero rRNA removal beads. The RNA is fragmented into small pieces and the cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and random primers, followed by second strand cDNA synthesis using DNA Polymerase I and RNase H. The resulting double-strand cDNA is used as the input to a standard Illumina library prep with end-repair, adapter ligation and PCR amplification being performed to produce a sequencing ready library. The final purified product is then quantitated by qPCR before cluster generation and sequencing.  The libraries were run on the Illumina HiSeq3000/4000 using HiSeq 3000/4000 SBS kit chemistry, 2x150bp paired-end sequencing run.