Place the Durham Tubes in the culture tubes.....aseptically add the broth and
carbohydrate in the volume desired. Put the tubes into an autoclave and run on
a liquid cycle for 5-10 minutes at 110 C. The heat will not have an ill effect
on the carbohydrates in the broth. Note, this time and temp is not sufficient
to sterilize the media, but it will get the trapped gas out of the tubes. At
ATCC, we autoclave the broth base, add filter sterilized carbohydrates,
dispense aseptically into sterile tubes (with inverted Durhams), and then run
the short liquid autoclave cycle to expel air.
Martin Gross
Supervisor of Media Preparation
American Type Culture Collection
