Whole genome sequence of Bacillus anthracis strain Kanchipuram
The paired-end sequencing library was prepared using Truseq Nano DNA Library prep kit. The library preparation process was initiated with 200 ng input. DNA was mechanically sheared into smaller fragments by covaris followed by continuous step of end-repair where an A is added to the 3 ends making the DNA fragments ready for adapter ligation. Following this step, platform-specific adapters are ligated to both ends of the DNA fragments. These adapters contain sequences essential for binding dual-barcoded libraries to a flow cell for sequencing, allowing for PCR amplification of adapter-ligated fragments, and binding standard Illumina sequencing primers. To ensure maximum yields from limited amounts of starting material, a high-fidelity amplification step was performed using HiFi PCR Master Mix. After obtaining the Qubit concentration for the library and the mean peak size from Bioanalyser profile, library were loaded onto illumina platform for cluster generation and sequencing. Paired-End sequencing allows the template fragments to be sequenced in both the forward and reverse directions. The library molecules will bind to complementary adapter oligos on paired-end flow cell. The adapters are designed to allow selective cleavage of the forward strands after re-synthesis of the reverse strand during sequencing. The copied reverse strand is then used to sequence from the opposite end of the fragment.
SUB7902252
BA