Targeting rate at the locus Tsg101 in MelJuSo cells
MelJuSo cells were grown in RPMI supplemented with 10% heat-inactivated fetal bovine serum (IFS) at 37C and 5% CO2. Genomic DNA in the vicinity of the sgRNA target was amplifie by PCR. Cells were lysed at 55C for 2 hours using tail lysis buffer (1% SDS, 100 mM NaCl, 50mM Tris, and 1mM EDTA) containing proteinase K (Roche Diagnostics; #03115879001). Genomic DNA was purified from the lysates by isopropanol precipitation. Libraries were prepared according to Illumina's instructions accompanying the DNA Sample Kit.