mm/dd/yyyy
PCR clean up of EsaI PCR and mCh Sender Intermediate for golden gate. Forgot to do DpnI digest of mCh (don't need to for EsaI because the PCR template is kan resistance).
| Sample
|
260
|
280
|
260/280
|
ng/µL
|
| mCh Sender int gg |
0.055 |
0.031 |
1.774 |
55.337
|
| EsaI |
0.046 |
0.026 |
1.776 |
45.692
|
picked all the colonies on the LuxR, RhlR, EsaR golden gate receivers with the 10:1 ratios and streaked them on a new plate. dotted 8ul of sender cells in the center of the plate. put at 37°C at 11am.
Going to run golden gate anyway. Backbone plate should have the same number of PCR template transformants as any of the ligation plates.
200fmol of insert, 20fmol backbone
| Reaction
|
Rec Bb
|
LuxI
|
BsmBI
|
T4 Ligase
|
T4 Ligase buffer
|
H2O
|
| Bb ctrl |
0.77 |
0 |
0.5 |
1.25 |
2 |
15.48
|
| RhlI |
0.77 |
3.2 |
0.5 |
1.25 |
2 |
12.28
|
| LasI |
0.77 |
2.8 |
0.5 |
1.25 |
2 |
12.68
|
| EsaI |
0.77 |
1.9 |
0.5 |
1.25 |
2 |
13.58
|
Set up PCR of LuxI with P131 and P132 AT 50°C ext time 1 min because it failed the other day with an annealing temp of 50.
| 
Project name
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