gRNA-seq of homo sapiens: breast cancer T47D cells
CRISPR screening gRNA-seq: About 270ug genomic DNA per group was amplified at 5 g per reaction (54 times) with specific primers using NEBNext Ultra II Q5 Hot Start High-Fidelity 2X Master Mix (NEB, #M0544). The presence of 162-base pair (bp) PCR products was verified using agarose gel electrophoresis. And then all the verified PCR products were pooled and purified by the TIANgel Maxi purification kit (TIANGEN, #DP210). Amplicon sequencing was then performed with an Illumina platform (Novogene Technology Co., Ltd., China).