WGS of Mycobacterium tuberculosis: Human sputum
The library preparation was performed by following plexWell WGS-24 Library Preparation protocol (Part No. PWP24). Briefly, 5 ng of Qubit quantified genomic DNA was tagmented (fragmented and adaptor tagged) and uniquely barcoded with Index i7 simultaneously using Sample-Barcoding Plate (SBP24), set-up provided in the plexWell Kit. The uniquely barcoded index i7 DNA was pooled with other WGS samples in a 24-plex and 8-plex format, for further pool barcoding process using index i5 barcode reaction setup and subjected to 12 cycles of FILL-AMP PCR (72 C for 10 mins followed by denaturation at 95 C for 3 min,cycling (98 C for 30sec, 64 C for 15sec, 72 C for 30sec) and 72 C for 3mins) to enrich the adapter-tagged barcoded fragments. The PCR product was purified using MAGwise Beads (MG5000).