IGEM:Virginia/2012/Notebook/Genetically engineered bacteriophage for diagnosis of whooping cough/2012/09/02

Week of 9/2/12

Performed pregnancy test experiment again: some cells were not lysed with B-PER along with a sample of 3:1 cell to B-PER ratio allowed to wait for several days at 4°C, resulting in the expected negative results for the former and faint, yet visible bands for the latter. The experiment will be redone to ensure accuracy of result making sure to spin down the cells and remove the LB media before lysing.
Transformed DH5α with BBa_K331022 again on an ampicillin plate and incubated overnight at 37°C.
Performed SynCG (with X+P), T7 (with BclI), backbone (with E+S), and backbone (with E+P) digests. The SynCG failed to form a band on the gel. The T7 could not be separated into small and large fragments. However, the BB fragments fortunately digested correctly.
Digested and nanodropped both successful backbones along with the digested RFP (with X+P), SynCG (with X+P), hCG (with E+S), bbp9 (E+S), bbp11 (E+S), T7 (with BclI), and some previously purified promotor and RBS.
Ligated the following combinations:
o bbp9 and backbone (E+S)
o bbp11 and backbone (E+S)
o hCG and backbone (E+S)
o promotor RBS, RFP, backbone (E+P)
o promotor RBS, SynCG, and backbone (E+P)