MONDAY, JULY 13TH
On Monday, the Enzyme Team was waiting on our dNTPs to arrive in order to PCR, but they never did. We used this time to further develop our project theme of pirates! Kill Switch team mini prepped and inoculated liquid cultures.
TUESDAY, JULY 14TH
On Tuesday, dNTPs had still not arrived, but the Enzyme Team was able to do a yeast transformation on Histine plates. We also continued our idea to make a prototype of a bioreactor by creating a prototype team and set up a meeting with Dr. Mosier. Kill switch team autoclaved and worked on developing our wiki.
WEDNESDAY, JULY 15TH
On Wednesday, our DNTPs finally arrived so we were able to PCR. Because our Gibson assembly did not work, we looked into PCR overlap, which would connect the two G blocks by using PCR. We PCR amplified, and ran a gel for manganese peroxidase, the first three wells being without primers, and the second three with. One of the bands with primers appeared, so the next day, we planned to run the rest of the manganese peroxidase. Kill switch team worked on mini-prep, restriction digest, and ligation.
THURSDAY, JULY 16TH
On Thursday morning, we PCR amplified lignin peroxidase, gel extracted the manganese peroxide from the day before, and ran another gel of the rest of the manganese peroxide. The bands from the second gel were not in the correct place, so we did not gel extract. This may have been caused from problems with sterile technique or with the PCR overlap, so we decided to do another PCR overlap of lignin peroxidase. Kill switch team did transformations.
FRIDAY, JULY 17TH
On Friday morning, we ran a gel of lignin peroxidase in which nothing appeared. We looked into optimizing our PCR for PCR overlap and anything that might make something work. Enzyme team ended the day with more PCR and Kill Switch team was in “depression because nothing worked”.- Kate