GSM5020527: hA3FBE3:T1,E1,FF,S3,S4 rep3,R2 rep2; Homo sapiens; OTHER
Target genomic sequences were PCR amplified by high-fidelity DNA polymerase PrimeSTAR HS (Clonetech) with primer sets flanking examined sgRNA target sites. For deep target genimoc DNA sequencing, Indexed DNA libraries were prepared by using the TruSeq ChIP Sample Preparation Kit (Illumina) with minor modifications. Briefly, the PCR products amplified from genomic DNA regions were fragmented by Covaris S220, and then PCR amplified by using the TruSeq ChIP Sample Preparation Kit (Illumina). After quantitated with Qubit High-Sensitivity DNA kit (Invitrogen), PCR products with different tags were pooled together for deep sequencing by using the Illumina HiSeq X10 (2×150) at CAS-MPG Partner Institute for Computational Biology Omics Core, Shanghai, China.