Sterilize the inoculating needle in the blue flame of the bunsen burner till red hot and then allowed to cool.
From the rack, take the Trypticase soy broth tube containing the 24-48 hour culture, remove the cap and flame the neck of the tube.
Using aseptic technique, take the culture of the organism from the TSB (tryptic soy broth) tube with the needle.
Again flame the neck of the tube and replace the tube in the test tube rack.
Take a sterile TSI slant tube from the rack, remove the cap and flame the neck of the tube.
Stab the needle containing the pure culture into the medium, upto the butt of the TSI tube, and then streak the needle back and forth along the surface of the slant.
Again flame the neck of the TSI tube, cap it and place it in the test tube rack.
Incubate at 37oc for 18 to 24 hours.