Whole genome sequencing of Mycobacterium leprae strain Br1
DNA was extracted from 101 human skin biopsies with known BI using a customized in-house protocol combining host tissue digestion and the QIAmp microbiome kit for host DNA depletion, strong bacterial cell lysis and silica based purification. Punch biopsies (6 mm) in 70% ethanol were first rehydrated in Hank balanced solution prior to mincing with scissors. Cells were detached from the tissue by 30 min incubation at 37°C with a mixture of 0.5 U of collagenase and dispase, followed by incubation at 56°C with 10 mg/ml of trypsin until complete digestion. Free cells were then suspended in 1 mL of phosphate-buffered saline (PBS) and DNA was extracted using the QIAmp DNA microbiome extraction kit according to the manufacturer recommendations. Each run of extraction included a batch of 5 to 9 samples and one blank control (500 ul of Hank balanced solution). Libriaires were prepared using the Kappa HyperPrep kit.