DNA double strand break mapping in AML patients: Sample KMT2A_1
Genomic DNA was purified by gently lysing cells in 50 mM Tris–HCl (pH 8.0), 100 mM EDTA, 100 mM NaCl, 1% SDS, 1 mg/mL Proteinase K for 3 h at 55_C followed by organic extraction purification and ethanol precipitation. Purified genomic DNA was subjected to blunting/A-tailing reactions, Illumina P5 adaptor ligation to capture broken DNA ends. Excess adaptor was removed and then DNA was fragmented by sonication, and subsequently ligated to Illumina P7 adaptor, and the libraries were PCR-amplified for 15 cycles. Prepared libraries were then subjected to whole-genome paired-end sequencing with the Illumina platforms.