title: GSM8549759: evoCDA, replicate 1, CD19 negative, pooled, endogenous; Homo sapiens; OTHER
library_construction_protocol: Genomic DNA was isolated using DNeasy Blood & Tissue Kit (Qiagen). For sgRNA count, the sgRNA cassette was PCR amplified from genomic DNA using primers carrying partial Illumina adaptor. PCR products were then re-amplified using primers carrying sample-specific barcode combinations. The endogenous CD19 enhancer sequence was PCR-amplified and barcoded using a similar strategy. Distributions of edits within a 346 bp fragment of the CD19 enhancer were obtained by targeted amplicon sequencing using MiSeq at PE250 and PE300 configuration. Guide RNAs were enumerated by NGS as previously reported (Seruggia et al. Mol Cell 2019).