GSM8506755: HCT-15 cells, WT, biol rep 3; Homo sapiens; RNA-Seq
Both knockout clones and wild-type cells from three different passages were detached using Versen solution, washed with PBS, and the resulting cell pellets were stored at −70 °C until further analysis. RNA extraction was performed immediately before the preparation of sequencing libraries using QIAGEN RNeasy Kit (Qiagen) following the manufacturer's protocol. RNA Integrity Number (RIN) was measured using Agilent 2100 bioanalyzer. Agilent RNA 6000 Nano or Qubit RNA Assay Kits were used to measure RNA concentration. KAPA RNA Hyper with RiboErase (KAPA Biosystem) Kit was used for further depletion of ribosomal RNA and library preparation. Different adaptors were used for multiplexing samples in one sequencing run. Library concentrations and quality were measured using Qubit ds DNA HS Assay kit (Life Technologies) and Agilent Tapestation (Agilent).