GSM9229278: Cal33, Metformin, rep3; Homo sapiens; OTHER
Genomic DNA was extracted and purified using a QIAGEN DNeasy Blood & Tissue kit. The human Brunello whole-genome CRISPR pooled library was a gift from David Root and John Doench (Addgene #73178). The library contains 76,441 sgRNAs targeting 19,114 genes (four sgRNAs per gene) and 1000 nontargeting sgRNAs as the negative control. The library DNA was transformed by electroporation using electrocompetent cells. The entirety of isolated genomic DNA was used for subsequent amplification through PCR to ensure capturing the full representation of the libraries. PCR products were sequenced on a HiSeq4000 instrument.