sgRNA sequencing library of HeLa HMGCR-Clover RNF145 KO, unsorted
DNA was extracted from  6e6 cells pre-sort using the Gentra Puregene Core kit A (Qiagen). Individual integrated gRNA sequences were amplified by PCR (primers: sgRNA_outer_F: GCTTACCGTAACTTGAAAGTATTTCG; sgRNA_outer_R: CGAGACTAGTGAGACGTGCTAC). Illumina adaptors  were added in an additional round of PCR (P5-sgRNA_inner_F: AATGATACGGCGACCACCGAGATCTACACTCTCTTGTGGAAAGGACGAAACA CCG; P7-index-sgRNA_inner_R: CAAGCAGAAGACGGCATACGAGATnnnnnnnGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTCCATTTGTCACGTCCTGCACG. Sequencing was carried out using the Illumina MiniSeq platform (Illumina sequencing primer (5'-3'): CTTGTGGAAAGGACGAAACACCG.