GSM7051218: siRNA-RP0581-1 subject 97265; Mus musculus; RNA-Seq
Approximately 10 mg of each liver tissue sample was homogenised in 0.55 mL phenol/guanidine-based QIAzol Lysis Reagent (Qiagen, 79306) using a Tissuelyser II (Qiagen). Liver tissue stored in RNAlater was used for primary analysis. The homogenate was extracted with 1-bromo-3-chloropropane (Sigma-Aldrich, B62404). RNA was extracted from 0.2 mL of the aqueous phase using the MagMax Technology (ThermoFisher Scientific) according to the manufacturer's protocol. RNA was quantified using spectrometry at 260 and 280 nm. 10 ng - 1 µg purified RNA from each sample was used to generate a cDNA library using the NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina. The cDNA library was then sequenced on a NextSeq 500 using NextSeq 500/550 High Output Kit V2 (Illumina).  Sequence reads were again aligned using STAR against the Ensembl mm10 transcriptome annotation. NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina