title: sgRNA sequencing library of HeLa HMGCR-Clover, sorted
design: Using the Gentra Puregene Core kit A (Qiagen), DNA was extracted from  5e6 cells sorted on HMGCR-Clover(high) after overnight sterol depletion and sterol repletion for 5h. Individual integrated gRNA sequences were amplified by PCR (primers: sgRNA_outer_F: AGGCTTGGATTTCTATAACTTCGTATAGCATACATTATAC; sgRNA_outer_R: ACATGCATGGCGGTAATACGGTTATC). Illumina adaptors  were added in an additional round of PCR (P5-sgRNA_inner_F: AATGATACGGCGACCACCGAGATCTACACTCTCTTGTGGAAAGGACGAAACACCG; P7-index-sgRNA_inner_R: CAAGCAGAAGACGGCATACGAGATnnnnnnnGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCCGACTCGGTGCCACTTTTTC. Sequencing was carried out using the Illumina HiSeq platform (Illumina sequencing primer (5'-3'): AGACTATAAGTATCCCTTGGAGAACCACCTTGTTGG .