title: GSM9051439: GPX1-eGFP clone#10 dim population; Homo sapiens; OTHER
library_construction_protocol: Genomic DNA was isolated from unsorted cells using the MasterPure Complete DNA Purification kit (Lucigen). Genomic DNA from sorted cells was isolated by phenol-chlorofom extraction. CRISPR screen custom DNA library. Sequencing libraries were generated using two sequential rounds of PCR using Herculase II Fusion DNA polymerase (Agilent). In the first round of PCR, 5µg of genomic DNA from unsorted cells was used in each of 24 100 µl reactions, and all recovered genomic DNA from sorted cells was split into two 100 µl reactions. Following 18 cycles of PCR1 amplification, 5 µl from the pooled reaction product was used in the second round of PCR (10 cycles), using primers containing barcodes and Illumina sequencing adaptors.