title: GSM5511207: K562-Lib7-Rep2; Homo sapiens; OTHER
library_construction_protocol: Genomic DNA (gDNA) was isolated using DNeasy Blood & Tissue Kits according to the manufacturer's protocol (Qiagen). The region that was integrated with genomic DNA including both the gRNA, target sequence, and barcode was PCR amplified from genomic DNA for HTS. Primers for HTS were used to specifically amplify all of the gDNA using 25 µl NEBNext Ultra II Q5 Master Mix (New England Biolabs), 17 µl of gDNA (~2.5ug), 2.5 µl of primer mix at a final concentration of 250 nM, and 5.5 µl water. For each gDNA sample, PCR reactions were in the range from 30 to 60.