JULY
Monday, June 30 - Friday, July 4
Assembled PFL 2 promoter and amplified via PCR. Ligated PLR promoter and attempted transformation with PAR promoter. Plated PAR promoter containing competent cells, but nothing grew. Plated again and still nothing grew. Ran restriction enzyme digest for PFL promoter. Ran ligation on PFL promoter and restriction enzyme digest on PFL2 promoter.
Monday, July 7 - Friday, July 11
Ligated for PFL 2 promoter. Ran a transformation for all four promoters; PLR, PAR, PFL, PFL2. Plates did not show sign of growth until Saturday.
Monday, July 14 - Friday, July 18
PLR, PFL, and PFL2 had cultures grow on their respective plates. PAR had no cultures grow. Made overnight cultures from PFL, PFL2, PLR and ran a colony PCR from the same samples we used for the overnight cultures. Ran our PCR products through a 0.7% agarose gel and no bands were present. Ran another colony PCR with PFL and PLR while incubating PFL2 to allow it to grow. Gel with PFL and PLR showed no bands.
Monday, July 21 - Friday, July 25
Miniprepped 4 samples; two from PFL and two from PLR. Amplified extracted DNA via PCR. Gel with PCR products showed no bands. Made a new set of overnight cultures from the same plates as before, but different colonies.
Monday, July 28 - Friday, August 1
Ran a restriction enzyme digest on Savannah's miniprep products. Made 1X TAE. Ran a gel with restriction enzyme digest products, no bands appeared. Nanodropped our miniprep results to determine concentration of DNA. Selected one sample from each promoter that had an adequate amount of DNA and ran a gel, which showed no bands. Plated PAR in order to culture cells, which plates showed no sign of later.
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