Module 1, Day 3

Transformation Efficiency

(a) [math]\displaystyle{ (10^9 cfu/ug) (\frac{1ug}{1000ng}) (\frac{1}{1000}) = 1000 colony forming units }[/math]
(b) [math]\displaystyle{ (50cfu)(\frac{1ug}{10^9cfu}) (\frac{100}{1}) (\frac{1}{2ul}) = 2.5 x 10^{-6} ug/ul }[/math]

Transformation Outcomes

Outcome 1: No bacteria were able to grow on the +kan plates, suggesting that none of the plasmids, including plate 1 with the positive control, were successfully transformed.
Outcome 2: The plates was either not treated with kanamycin, or a contamination occurred.
Outcome 3: The kill-cut mechanism failed, and the sticky ends of the backbones transformed into plate 3 were able to come together despite our efforts.

Plasmid Map

See Attached Sheet

Plaque Assay Titer

[math]\displaystyle{ Titer = (\frac{PFU}{Dilution * Volumes}) }[/math]

• K01:

     [math]\displaystyle{  Titer = \frac {880}{10^{-6}*10ul*\frac{1ml}{1000ul}} = 8.8 x 10^{10} pfu/ml }[/math]

• E4:

     As detailed in my lab notebook, we failed to observe E4 pfu colonies on the lb-plates, most likely due to  
     uneven spreading and bubbles in the agar. As a result, I do not have the data available to calculate the 
     titer for E4. 

It makes sense that K07 would form less PFUs than E4--as a helper phage to other strains of M13, it is probably less effective at infecting bacteria with its own genome. Curiously however, our K07 colonies proliferated in similar number and morphology to other groups' E4 colonies.

Refactoring Bacteriophage T7

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