title: GSM8890254: HepG2 siDDX55 replicate 2 QuantSeq Rev V2 3'end seq of whole cell RNA; Homo sapiens; RNA-Seq
library_construction_protocol: Cells were lysed on ice for 5 minutes in 10mM Tris-HCl pH 8.0, 0.32 sucrose, 3mM CaCl2,  2mM MgCl2, 0.1mM EDTA 0.1% NP40 1mM DTT plus 0.04U/ul RNasin. Total RNA was extracted using TRIzol following the manufacturer's protocol. For fractionation samples, low speed centrifugation was used to separate the fractions with the supernatant added to TRIzol for cytoplasmic and pellet lysed in TRIzol for nuclear fractions. RNA quality for all samples and fractions was verified using a BioAnlyzer. Further QC and library preparation using QuantSeq Rev V2 (Lexogen) was carried out by Lexogen and subjected to single end, stranded sequencing.