Survival test

This experiment will test to see if our modified organisms will survive in different conditions. One part is to see if our bacteria holding the plasmid with the gene will be outcompeted by a natural bacteria, and one test is to see if the bacteria will simply lose the plasmid with time. We will compare these two results to see if our bacteria containing our plasmids will actually survive in a environment without selective pressure for the plasmid.

Natural bacteria VS modified bacteria with plasmid

1. *Grow overnight culture, 1 ml of natural and modified bacteria in LB with the modified and
natural bacteria separated from eachother. Use no antibiotics in the media.
*Also grow 1 ml of each of the modified bacteria mixed with the natural.

2. *The next day, take 500 µl of the separated cultures and mix together by vortexing. Take 1 µl
of this and grow in 1 ml lb medium overnight again.
*Also plate the overnight culture with the bacterial mix from yesterday on both resistance
plates and non resistance plates.
*Also plate each of the separated bacterial cultures from yesterday.
*Also take 1 µl of each of the separated cultures from yesterday and grow again in 5 ml of
media.

3. Repeat all the steps again and plate the different cultures on resistance plates and the
natural as control on resistance free plates.