WGS of Mycobacterium tuberculosis: adult sputum
Sequencing library was generated using MGIEasy Universal DNA Library Prep Kit V1.0 following manufacturers recommendations and index codes were added to each sample. Briefly, genomic DNA sample was fragmented by sonication to a size of 350 bp. Then DNA fragments were endpolished, A-tailed, and ligated with the full-length adapter for BGI sequencing, followed by further PCR amplification. The constructed library was qualified with an Qubit 4.0 Fluorometer (ThermoFisher, Foster City, CA, US). And the library fragment size was analyzed with Qsep-100 (Hangzhou Houze Biotechnology Co., Ltd.). Then use Library Quant Kit (kapa,Roche, Rotkreuz, Switzerland) to accurately quantify the library for screening qualified libraries. The qualified library was pooled at 3nmol each. After circularization and generating DNBs, the resulting libraries were sequenced with MGI High-throughput Sequencing Set (PE150) on MGISEQ-2000/T7 platforms (MGI, Shenzhen, China).