Field of research:
Lab: Jill Bargonetti
Manhattan Hunter Science High School
10th/ 11th Grade
CLONING AND CONSTRUCTING EXPRESSION PLASMIDS WITH shRNA INSERTS
This summer, I completed an internship at Dr. Jill Bargonetti's lab. My
project there was to clone and construct plasmids with shRNA inserts.
These plasmids will be used in future experiments for silencing p53,
mdm2 and mdmx gene experession in breast cancer cells. We began with
two different types of plasmids: psM2c (contains shRNA) and STGM
plasmids (inducible promoter). We digested both with Xho1 and EcoR1
restriction enzymes for specific ligation. Then we ligated the shRNA
and expression plasmid by using ligase. Next, we introduced the
recombinant plasmid back into the bacterial cell and cloneD them.
Finally, we purified the plasmid by following miniprep protocol. These
plasmids were then stored in the refrigerator at -20 degrees Celcius
until the next experiment.
Working in a big lab like this is very hard. It takes a while to understand what people are doing in the lab. Once you are able to understand, it's fun.