title: ASCC3 promotes chemosensitivity in colorectal cancer cells
design: ASCC3 promotes chemosensitivity in colorectal cancer cells
library_construction_protocol: HCT116 parental (WT) and ASCC3-KO cells were maintained at 37°C with 5% CO₂ in DMEM supplemented with 10% fetal bovine serum, L-glutamine, non-essential amino acids, 100 U/mL penicillin and 0.1 mg/mL streptomycin; cultures were routinely screened for mycoplasma by DAPI staining. Cells were harvested for RNA extraction. Total RNA was extracted with TRIzol and purified using the RNeasy Mini Kit (Qiagen, cat. no. 74106) per manufacturer's instructions; RNA integrity was confirmed by BioAnalyzer. Libraries were prepared using the NEBNext® Ultra™ II Directional RNA Library Prep Kit (New England Biolabs) with poly(A) selection, following the manufacturer's protocol.