GSM9207629: MTHFD1-K56R cells cultured in dialysed IMDM supplemented with 50µM adenosine, replicate 2; Homo sapiens; OTHER
After the end of the genome-wide KO screen, remaining cells were collected and gDNA was isolated using the DNeasy Blood and Tissue Kit (Qiagen). The library was amplified using Takara Ex Taq DNA polymerase, yielding the sgRNA sequence. P5 and P7 index primers in unique combination were used for amplification (primers adapted from Mol Cell 75, 849-858 e848 (2019)). PCRs were run with cycle numbers below saturation, as judged by initial control PCRs. Gel bands of expected size were gel extracted and gel purified. Quantity was determined using Qubit and equal amounts were pooled and sent for sequencing.