title: HK-2 cell treated  with vancomycin and detected by CRISPR/Cas9 library screeningbiological replicate 3
design: Genomic DNA was extracted using the Quick-gDNA MidiPrep (Zymo Research, cat. no. D3100) according to the manufacturers protocol and concentration was measured on the Nanodrop (Thermo). PCR was carried out to amplify the sgRNA for NGS. 2.5 ug of gDNA were arranged in 50ul reaction mix for each of the 10 Diversity-Fwd primers and 6 Rev-barcode primer. Each PCR reaction consisted of NEB Next High Fidelity PCR Master, gDNA, Diversity-Fwd primers and 6 Rev-barcode. Pooled PCR products were purified with the Zymo-Spin V with Reservoir kit (Zymo Research, cat. no. C1016-25) according to the manufacturers instruction.