GSM7696491: HeLa cell, Day 30, replicate1; Homo sapiens; OTHER
The genomic DNA was isolated from 4 × 106 cells using the Genomic DNA Kit (TianGen Biotech) for each replicated sample, and a total of 32 μg of the recovered products, representing at least 200 × coverage of the initial library size, were used as amplification templates in 16 independent PCR tubes (50 μl system each). The genomic barcode regions encoding paired crRNAs introduced by lentivirus infection were PCR amplified using Q5 High Fidelity Polymerase (NEB) with 22 cycles using two primers targeting the upstream U6 promoter and the downstream CMV promoter. In a second round of PCR, adaptors for sequencing purposes with different barcodes were added for different samples (replicates and time points). Finally, the PCR products of all samples were pooled and purified with DNA Clean & Concentrator-5 kit (Zymo Research).