Bacillus Calmette-Guérin (BCG) Vaccine Production

Begin by collecting the BCG culture at the mid or late log growth phase. Centrifuge it had 3000 G for 10 minutes, then discard the supernatant. Next, add 10 milliliters of PBS to wash the bacteria.

After centrifugation, discard the supernatant. Resuspend the pellet in five milliliters of infection media. Place the tube in an ultrasound bath for 15 minutes at full power with a frequency of 80 hertz.

Now centrifuge at 1, 000 G for 10 minutes. Collect the supernatant and discard the pellet. Measure the optical density of the supernatant.