After 24 hours, growth was as expected on all 4 plates.
1) LB -pGLO
This plate showed growth, indicating that our conditions were right for E. Coli to grow, eliminating environment as a possible cause of failure, should that have occurred. The cells were not responsive to UV-light, as expected.
2) LB/amp -pGLO
This plate showed no growth, as the cells on that plate had not taken up the pGLO plasmid and therefore were not resistant to the ampicillin present in the agar gel.
3) LB/amp +pGLO
This plate showed growth, indicating successful absorption and integration of the pGLO plasmid into the DNA sequence of the E. Coli, specifically through resistance to the ampicillin present in the agar gel. The cells were not responsive to UV-light due to the absence of arabinose, as expected.
4) LB/amp/arab -pGLO
This plate showed growth, indicating successful absorption and integration of the pGLO plasmid into the DNA sequence of the E. Coli, both through resistance to the ampicillin present in the agar gel and the responsiveness of the cells to UV-light, emitting a greenish glow. Due to the presence of the arabinose in the agar gel, the gene coding for GFP production was turned on, allowing for GFP to be made.
Transformation Efficiency
Total # of cells growing on agar
plate = transformation efficiency
Amount of DNA spread on agar plate
1 loop = ~10μL
[pGLO] = 0.08 μg/μL
10μL = 0.8μg pGLO
250μL CaCl2 + 250μL LB Broth + 10μL pGLO = 510μL total liquid
120μL +pGLO suspension plated ~
20%
510μL total liquid
.08μg pGLO x .20 = .016μg pGLO transformed cells
610 cells growing = 3.8 x 103
= transformation efficiency
.016μg pGLO