This project includes Ion Torrent whole genome sequencing of Burkholderia pseudomallei isolates obtained from clinical samples of melioidosis patients at AIIMS Bhopal, Madhya Pradesh, India.
The whole genome libraries were prepared as per the manufacturers (Thermofisher)instructions. Briefly, the Ion Plus Fragment DNA Library Prep Kit (Thermofisher, Cat. No.4471269) was used. Genomic DNA was first sheared to ~250 bp fragments using the IonShear Plus Kit , followed by adapter ligation and barcode tagging with the Ion XpressBarcode and Adapters Kit (Thermofisher, Cat. No. 4471250). After each step, purificationwas performed using AMPure beads (Beckman Coulter, Cat. No. A63881). Library size isdetermined by using 2% agarose gel size select (Thermofisher, Cat. No. G661012).Library quantification was performed using the Qubit High-Sensitivity kit. The libraries werethen diluted to a final concentration of 100pM and subjected to cluster amplification. Thelibraries were loaded onto 540 chip using the Ion Chef System (Thermofisher, Cat. No.2664777) and sequenced on the Ion GeneStudio S5 Plus Series System forcomprehensive Burkholderia pseudomallei genome sequencing. The raw sequencing dataobtained from the Ion GeneStudio S5 Plus Series System were further processed andanalyzed.