After finishing the presentations, we used DNA from our bacteria to run a restriction enzyme digest. This was done to determine if our gene transfer was successful. We started by adding 23 µL of sterile water to three microfuge tubes. We then added three µL of buffer, three µL of the DNA, and one µL of the appropriate restriction enzyme (either NheI or EcoRI). For the third tube, we only used 22 µL of water but we added one µL of both NheI and EcoRI. This will allow us to be certain that the gene transfer worked. After allowing the enzymatic reaction to work during lunch, we added six µL of dye (in order to reach the correct concentration of dye) and loaded the digests into electrophoresis wells.
Here is the gel that was run:
Below is a DNA ladder that will help with our analysis:
After loading the electrophoresis chambers, we took single bacterial colonies off of our original Petri dishes and streaked new plates. We then placed the new plates in the incubator and disposed of the old plates.