Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2015/02/18

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02/18/2015

Ligation results:

Plate # colonies size of colonies
+ ctrl lawn tiny
- ctrl 26 large
Bb 50ng 37 large
Bb 100ng 32 large
1:1 21 large
1:3 32 large
3:1 27 large

The ligation resulted in colonies, but the negative control, which did not have any plasmid DNA, had colonies as well—this was a concern.
The reasons why the negative control could have grown colonies include:

  • The ampicillin in the plates broke down, allowing other bacteria to get in and grow.
  • Some plasmid DNA got into the negative control while pipetting

To test and see if it was the plates, Ben created a negative control without DNA and incubated the plate overnight.
If the plates were the problem, we would have seen colonies on his plate as well, but this was not the case. <br Since we are not 100% certain what the problem is, we decided to pick colonies and test them to see if they are what we hoped them to be.