GSM8080319: gwscreen_JS056_NUP98_KDM5A; Mus musculus; OTHER
Genome-wide CRISPR screen: Genomic DNA was isolated by cell lysis (10 mM Tris-HCl, 150 mM NaCl, 10 mM EDTA, 0.1% SDS), proteinase K treatment and DNAse-free RNAse digest, followed by two rounds of phenol extraction and 2-propanol precipitation. Genome-wide CRISPR screen: Sequencing libraries were generated using a two-step PCR protocol. The first PCR was performed using 0.2 μl of AmpliTaq Gold in 50-μl reactions containing 1 μg of genomic DNA. For each sample, the resulting PCR products were pooled and purified using MBSpure magnetic PCR purification beads and used as input for a second PCR introducing the standard Illumina adapters using 10 ng of DNA template.