title: GSM8163306: Liver, LeuD, WT, 03 bio rep1; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was isolated using the Direct-zol™ RNA MiniPrep Plus Kit (Zymo Research) following instructions provided by the manufacturer. RNA sequencing libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit for Illumina (New England Biolabs) following the manufacturer's instructions. Briefly, mRNAs were initially enriched with Oligod(T) beads. Enriched mRNAs were fragmented for 15 minutes at 94 °C. First strand and second strand cDNA were subsequently synthesized. cDNA fragments were end-repaired and adenylated at 3'-ends and universal adapters were ligated to cDNA fragments, followed by index addition and library enrichment by PCR with limited cycles. The sequencing libraries were validated with the TapeStation System (Agilent Technologies) and quantified by using Qubit 2.0 Fluorometer (ThermoFisher Scientific). Further validation of the libraries was carried out by quantitative PCR (KAPA Biosystems) of selected genes.