sgRNA sequencing of DSB-Spectrum screen:Clone D9_n2 and n3
HEK 293T cells were infected to integrate a custom-generated sgRNA library with 12018 different sgRNAs. The genomic DNA was extracted, and sgRNA sequences were amplified, followed by PCR purification. Next , a second PCR step was performed to add the sequencing primer binding site, the illumina adapters, and the indeces. Eleven samples were loaded on one lane, these are the indeces per sample: D9_Total1_n2 (TTGATATT), D9_BFP- (CGAGGCCT), D9_GFP_n2 (TGAGCAAT), D9_Total2_n2 (TGTTAATG), D9_BFP+_n2 (TTCGTTGG), D9_Total1_n3 (GCCACCGG), D9_BFP-_n3 (ATGAACCG), D9_GFP+_n3 (ATCGCCAG), D9_Total2_n3 (TCGGGCAG), D9_BFP+_n3 (CGAAGTTC), Library_n2 (TCAAACAC).