Week 3 : Sink or Swim?

Some experiments are working better than others.

We are still having troubles with getting PCR to work, we tried diluting the samples even more and when that was unsuccessful we even proceeded to do a re-extraction. After the re-extraction the samples looked a lot cleaner and running them on a gel and nano drop confirmed that there was DNA in the samples. Unfortunately, in the PCR for Katablepharis none of the samples amplified except for a very faint band in the positive sample (which is unusual). We suspect that it could be the enzyme, and we just got a new stock of the phusion so perhaps it will give us better results.

Unlike PCR our cloning experiments have been going well. We successfully cloned some positive PCR samples into a vector that we inserted inside E.coli cells. Our cells were able to grow on ampicillin plates meaning they had the vector inside of them, many colonies were white indicating that the vector had and insert that disrupted the lacZ gene. To check that the insert was in the vector we cut the vector with restriction enzyme and most of our samples had the vector in it so we were able to send the samples for sequencing. We will get the results next week.