GSM8683009: Huh7.5.1 Cas9-P2A-hygromycin cells, DENV2 replicon cells, d19, high-GFP sorted, replicate 2; Homo sapiens; OTHER
Genomic DNA was extracted from the cell pellets of the sorted cells using the Zymo Quick-DNA Miniprep Plus kit and from cell pellets of the unsorted cells using the Qiagen Blood Maxi Kit. The genomic DNA preps were further purified from contaminants using the Zymo OneStep PCR Inhibitor Removal Kit. Sequencing libraries were prepared from extracted genomic DNA by PCR amplification using barcoded Illumina P7 and P5 primers of the sgRNA cassettes. The entire prep of extracted gDNA was barcoded and amplified (1 µg gDNA/ 50 µl PCR reaction) for each screen condition.