For my last week Ben’s undergraduate student
came into lab. Her name is Karen and she is an undergrad at Penn. We had a lot
to do so Ben had me helping to train Karen with certain things, like running
PCR and HRM. Having Karen allowed for things to go much faster, and we got a
lot done. In order to check our Nog1-CRISPR mutants we did a DNA precipitation.
This process required us to “wash” the DNA by taking the liquid in which the
DNA pellet was suspended and adding ethanol, and then other alcohols. These
washes were done in order to break down free floating nucleotides and proteins
that could cause our gel to be messy.
The first gel we ran.
After this precipitation we ran the gel.
Running the gel started with having to make the actual gel, and that included
measuring out Agarose and 5xTBE buffer. To get the gel hot enough we had to
microwave it, and then pour the mixture into the mold. Loading the gel took a
short amount of time because both Karen and I were working at the same time.
The results of the gel were not very promising, as bands that should have been
the largest were faint, and ones meant to have little DNA were very bright. In
order to deal with this we redid the precipitation. This was on my last day so
I did not get to see the results, but what we were looking for in a successful gel
was a split in the bands. If the bands had two bright bands they would be
mutants. That is because these two bands would show that the DNA was cut. If
only one bright band, the DNA would be wild type.
For
my last week my lab had a goodbye celebration for me, mixed with a birthday
celebration for a few of the lab members. There were four cakes, as the lab’s
tradition is that you make a cake for the person whose birthday is before
yours. I had an amazing time at my lab and was so sad to leave, but many of my
lab members promised to stay in touch.
The Birthday note written on the board.
The many really good cakes made.
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