WGS of Homo sapiens: U2OS MSH6 KO
Genomic DNA was extracted using QIAGEN Genomic DNA Preparation. Briefly, 4x106 cells were collected by trypsinization, washed twice in cold PBS and finally resuspended in 0.5mL ice-cold PBS1X. Cytoplasmic fraction was first extracted twice with buffer C1. Purified nuclear fraction was completely resuspended in buffer G2. 25mL Proteinase K was added and samples were incubated at 50C for 1hr, and loaded on pre-equilibrated Genomic-tip 20/G with buffer QBT. The column was then washed twice with buffer QC, and DNA was finally eluted using 2mL of buffer QF. Genomic DNA was precipitated using 0.7 volumes isopropanol and finally resuspended in 200mL 10mM Tris-HCl pH 8.