RobWarden:M1D3

Module 1, Day 3 For Next Time

Transformation Math

First Transformation:
[math]\displaystyle{ 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} }[/math]

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

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	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
Diagnostic digest 2
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: [math]\displaystyle{ 1327\ \mbox{plaques} \div 10^{-6}\ \mbox{dilution} = 1.3 \times 10^9 }[/math]

M13 Titer: [math]\displaystyle{ 276\ \mbox{plaques} \div 10^{-6}\ \mbox{dilution} = 2.8 \times 10^8 }[/math]

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.