GSM4729895: A375_dCas9-dMSK1-no-treatment-_re1; Homo sapiens; OTHER
Genomic DNA was extracted using the Qiagen Quick-gDNA midi kit (Qiagen, #13343) following manufacturer's instructions. To amplify the gRNAs, we used 8 μg of genomic DNA as a template across eight 100-μL PCR reactions using Q5 hot start polymerase (NEB, #M0493L). We carried out amplification according to the manufacturer's instructions, using 25 cycles at an annealing temperature of 60 ° C with indexed primers. We purified the amplified libraries with QIAquick Gel Extraction Kit (Qiagen, # 28706). Each sample was quantified after purification with the Qubit dsDNA High Sensitivity assay kit (Thermo Fisher, Q32854). Samples were pooled and sequenced using the MiSeq platform (Illumina).