GSM8758462: E16,cWT,rep2; Mus musculus; RNA-Seq
Total RNA was isolated using the Quick-RNA Miniprep Kit (Zymo Research) according to the manufacturer's protocol. 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, Ipswich, MA). Total RNA (375 ng) was treated with the rRNA Depletion module. The rRNA-depleted RNAs were then fragmented for 10 min, determined by the RIN (RNA Integrity Number) of input RNA as per protocol, and subjected to the first-strand cDNA synthesis dUTP mix. Samples undergo end repair and dA-Tailing step followed by ligation of NEBNext adapters. The libraries were PCR-amplified and checked for quality and quantity by TapeStation (Agilent). The samples were pooled and sequenced on the Illumina NovaSeq S4 Paired-end 150bp, according to manufacturer's recommended protocols.