Module 1, Day 3 For Next Time

Transformation Math

First Transformation:
$10^9\ \frac{\mbox{colony forming units}}{\mu g} * \frac{1\ \mu g}{10^3\ ng} * 10^{-3}\ \mbox{dilution} * 1\ ng = 10^3 \mbox{colonies}$

Second Transformation:
$50\ \mbox{colonies} * \frac{\mu g}{10^9\ \mbox{colony forming units}} = 5 \times 10^{-8}\ \mu g$
$\frac{5 \times 10^{-8}\ \mu g}{2\ \mu L} = 2.5 \times 10^{-8} \frac{\mu g}{\mu L}$

Control Interpretations

Ligation No Colonies Thousands of Colonies
Positive Control Bacteria not competent Worked Well
bkb -ligase bkb is linear bkb was not digested
bkb +ligase killcut worked killcut did not work
bkb + insert insert recreated BamH1 site insert was ligated into bkb

Diagnostic Digests

 Diagnostic digest 1 Diagnostic digest 2 plasmid with insert plasmid no insert Enzyme(s) used BamH1 & XhoI BamH1 & XhoI Buffer used 2 2 Temperature 37 C 37 C Predicted fragments 8701 5702 & 2967 Enzyme(s) used EcoRV & DraIII EcoRV & DraIII Buffer used 3 3 Temperature 37 C 37 C Predicted fragments 3501, 2831, & 2337 6332, & 2337

Phage Titer

Collected Data

Phage Quarter-Plate Count x4
E4 343 1327
M14 69 276

Analysis

E4 Titer: $1327\ \mbox{plaques} \div 10^{-6}\ \mbox{dilution} = 1.3 \times 10^9$

M13 Titer: $276\ \mbox{plaques} \div 10^{-6}\ \mbox{dilution} = 2.8 \times 10^8$

The E4 Titer was 4.6 times larger than the M13 Titer. The extra glutamic acids on the E4 Phage may allow it to diffuse faster and therefore infect more bacteria.