Illumina sequencing of cloned B. pseudomallei gDNA fragments that drove expression of a downstream gfp reporter after infection of J774 cells
Plasmid DNA was extracted from GFP positive clones after three passages in macrophages. DNA inserts were amplified by PCR using primers that annealed to the plasmid backbone flanking the inserts. The amplified PCR products were used directly as input for Illumina sequencing. Sequencing libraries were prepared using a NEB Next DNA library preparation kit. Sequencing was performed on an Illumina MiSeq flowcell generating 250 bp paired end reads.