title: GSM9159680: H460,shNTC,Day0; Homo sapiens; OTHER
library_construction_protocol: Genomic DNA was extracted from cells using Macherey Nagel Nucleospin Blood XL kit (Item number- 740950.50) sgRNA regions of library cassette were amplified using Phusion HiFi Polymerase Master Mix in two PCR steps. In PCR 1, to increase sequence diversity, libraries are amplified in two primer orientations named SET A and SET B. In SET A orientation, the protospacer is sequenced during read 1. In SET B orientation, the protospacer is sequenced during read 2. The orientation of the amplicon is determined during round 1 PCR. In PCR 2 unique primers are used to tag each sample for the purpose of disambiguating each sample as they are pooled together