title: Illumina HiSeq 4000 sequencing; mRNA-seq of HCT116 USP22 knockdown cells
design: mRNA-seq of HCT116 USP22 knockdown cells
library_construction_protocol: HCT116 were grown in McCoy's medium supplemented with 10% fetal bovine serum, 100 units/ml penicillin and 100 μg/ml streptomycin at 37°C and 5% CO2. The following siRNAs were used: NT5 (non-targetting control) from Dharmacon, cat. no D-001210-05 and USP22 also from Dharmacon, cat. no: MU-006072-01-0010. siRNA transfections were performed using Lipofectamine® RNAiMAX (Invitrogen) according to the manufacturer's instructions. RNA was extracted usiung QIAzol® Lysis Reagent (Qiagen) according to the manufacturer's protocol. Libraries were prepared using TruSeq RNA Library Preparation Kit v2 (Illumina®) after verifying the RNA integrity on an agarose gel. Library quality was assessed using an Agilent Bioanalyzer 2100