Whole genome seqquencing of Phocaeicola vulgatus NMBE-5
Bacterial genomic DNA was processed using Illumina DNA prep workflow. DNA libraries were constructed from 50ng genomic DNA within 2 working days. The protocol initiated with magnetic bead-based "Tagmentation '' on BLT(Bead Linked Transposomes), which resulted in similar-sized DNA fragments. Next, the tagmented DNA was thoroughly washed using Tagment Wash Buffer(TWB) and cleaned by placing it on a magnetic stand. Further, the tagmented DNA is amplified using limited cycle PCR by using Nextera DNA Unique Dual (UD) indicies. In this PCR, Enhanced PCR Mix(EPM) was used using Index 1(i7) and Index 2(i5) adapters. Later, the Index PCR was purified using SPB(Sample Purification Beads). Quantification of all the purified indexed samples was done using Qubit 4 and equal molar concentration of all the samples were made (4nM). Simultaneously, the samples were checked for the exact library size using Tapestation. Libraries were loaded onto a MiSeq reagent cartridge and clustered on the MiSeq System. Paired-end sequencing was performed using MiSeq v2 reagents and 2  150 bp reads. Sequencing generated 12 million reads passing filter, with a total run time of 32 hours