IGEM:Virginia/2012/Notebook/Genetically engineered bacteriophage for diagnosis of whooping cough/2012/06/24

(6/24 – 6/30)

Performed Gibson Assembly on Luc genome and hCG genome. We then performed digest on the Gibson Assembly products. The results were hard to decipher.
We tried various lysis techniques such as: BPER buffer, chloroform, and RIPA buffer.
Bought EPT and Clearblue pregnancy test kits. Their purpose was to detect for the presence of hCG. Unfortunately we got all negative results. We tried with E. coli first, but still got negative results meaning that it did not take up the hCG gene.
Tried additional sonication method for lysis.
Digest of phage genome and vector digest both worked. During PCR purification, the products were resuspended in too large a volume, thereby making them dilute.
The next step was to either concentrate them or run a larger volume of digested hCG plasmid in order to detect via traniluminator.