Whole genome sequencing of Mycobacterium tuberculosis
Genomic DNA was extracted for sequencing using the cetyltrimethylammonium bromide method of DNA purification. Each extracted DNA was quantified by a Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA, USA). After the library was diluted, an Agilent 2100 bioanalyzer system (Agilent Technologies, Palo Alto, CA, USA) was used to detect the inserted fragments of the library. The quantitative polymerase chain reaction method was used to accurately quantify the effective concentration of the library to guarantee quality of the library after the size of the inserted fragments met the expectation. According to the requirements of effective concentration and target off-line data volume, different libraries were pooled into a flow cell. cBot was used to generate clusters, which were then sequenced using the Illumina NovaSeq 6000 (Illumina, San Diego, CA, USA) high-throughput sequencing platform.