GSM8070812: MiaPaCa2_apilimod_14d; Homo sapiens; OTHER
Genomic DNA was isolated using the DNeasy blood and tissue kit (Qiagen) according to the manufacturer's protocol. For each condition, sgRNA was amplified from 50mg gDNA using Herculase II Fusion DNA Polymerase (Agilent Technologies), column purified using Select-a-Size DNA Clean & Concentrator kit (Zymo Research), and then gel-purified using 6% Novex TBE gel (Thermo), followed by isolation from the gels with Gel Breaker Tubes and Gel Filters (BioChain). The resulting PCR products then underwent end-repair and A-tail addition followed by New England Biolabs (NEB) adapter ligation. The final library was prepared by enriching adapter-ligated DNA fragments using 2x KAPA HiFi HotStart mix and NEB dual code barcode following the manufacter's protocol. The libraries were then sequenced on an Illumina NovaSeq 6000 (paired-end, 300 cycles).