GSM6067658: Wild type rep 3; Homo sapiens; RNA-Seq
CAL51 cells were trypsinised, and resuspended in a lysis solution containing 150 mM NaCl, 0.1% Triton X-100 and 50 mM Tris-HCl (pH 8) for 30 min at 4°C. Then, cells were centrifuged at 21,000 g for 15 min at 4°C to clarify lysates. Protein concentration was determined using a Bicinchoninic acid (BCA) assay. RNA was isolated from CAL51 cells using a miRNeast MiniKit (Qiagen) incorporating an on-column DNase digestion according to the manufacturer's instructions. Libraries were constructed using an Illumina Stranded mRNA preparation kit and sequenced using a NovaSeq platform by AGRF.