GSM4745680: VEH1; Mus musculus; RNA-Seq
TRIZOL reagent RNA was assessed for quality using the TapeStation (Agilent, Santa Clara, CA). Samples were prepared using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (catalog number E7760L) Ribo depletion Module NEBNext rRNA Human/Mouse/Rat (catalog number E6310X) and NEBNext Multiplex Oligos for Illumina Unique dual (catalog number E6440L) (NEB, Ipswitch, MA). Where 5ng of total RNA was ribosomal depleted using rRNA Depletion module. The rRNA-depleted RNA is then fragmented 7 minutes determined by RIN (RNA Integrity Number) of input RNA as per protocol, and copied into first strand cDNA using reverse transcriptase and dUTP mix. Samples undergo end repair and dA-Tailing step followed by ligation of NEBNext adapters. The products are purified and enriched 16 cycles by PCR to create the final cDNA library. Final libraries were checked for quality and quantity by TapeStation (Agilent) and qPCR using Kapa's library quantification kit for Illumina Sequencing platforms (catalog # KK4835) (Kapa Biosystems,Wilmington MA).