WGS of Mycobacterium tuberculosis -  H37Rv: Thioquinazolinone resistant mutant
gDNA was enzymatically fragmented for 40 min at 37 C, followed by end repair and A-tailing using HyperPrep End Repair and A-Tailing enzyme by incubating for 30 min at 65 C. Next, the fragments were subjected to T-tailed 7-nucleotide barcodes adapter ligation using unique adapters from KAPA Unique Dual-Indexed Adapter Kit (KAPA Biosystems #KR1736) and DNA ligase by incubating the mix at 20 C for 15 min. Ligation was followed by a cleanup step using 0.8x beads (KAPA Pure Beads; Roche #7983280001). Next, the library was PCR amplified using library amplification primer mix and KAPA HiFi HotStart Ready Mix. Amplicons in the size range of 250-400 bp were selected using 1x bead-based cleanup and subjected to 2x75 bp paired-end sequencing on an Illumina NextSeq 500 mid output 150 cycle kit (reads, ~150 M reads/run).
SUB15083570
15_S14_RM14