*2.5. Mycobacterial Genomic DNA Preparation for the Multiplex PCR Assay and for attR and attL DNA Regions PCR*

For isolation of DNA from BCGwt, BCG.HTI2auxo.int; 2 mL of mycobacterial culture was centrifuged at 5000 rpm for 10 min at room temperature. The pellet was resuspended in 200 μL of distilled water and heated at 95 ◦C for 20 min to inactivate and lyse bacterial cells. The sample was next centrifuged at a speed of 13,000× *g*. A total of 5 μL of supernatant was used for the amplification reaction. The commercial BCG strains were treated similarly, except in this case, 400 μL of the reconstituted freeze-dried flasks were used.
