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Project name
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mm/dd/yyyy
Purified PCR reactions 5 and 6 from yesterday (receiver backbone) using the gel extraction columns and the DNA clean and concentrator kit. Got really low yield, need to look into a better kit for this.
Will set up golden gate with the purified PCR backbones. Will gel extract the other ones and then run those tomorrow if this doesn't work. Will only do one of each because I don't have enough receiver backbone.
Description
|
ng/µL
|
length
|
fmol/ul
|
ul for 200 fmol
|
Rec bb |
31 |
2203 |
21 |
9.52
|
pLux-GFP |
47.148 |
1051 |
68 |
2.94
|
pRhl-GFP |
50.466 |
1051 |
72 |
2.78
|
pEsa-GFP |
32.1 |
1051 |
46 |
4.35
|
pLac-LuxR |
45.771 |
817 |
89 |
2.25
|
pLac-RhlR |
59.88 |
765 |
119 |
1.68
|
pLac-EsaR |
37.847 |
758 |
74 |
2.70
|
Reaction
|
Rec bb
|
pLux-GFP
|
pLac-LuxR
|
BsmBI
|
T4 ligase
|
T4 ligase buffer
|
H2O
|
Lux Rec |
9.52 |
2.94 |
2.25 |
0.5 |
1.25 |
2 |
1.54
|
Rhl Rec |
9.52 |
2.78 |
1.68 |
0.5 |
1.25 |
2 |
2.27
|
Esa Rec |
9.52 |
4.35 |
2.7 |
0.5 |
1.25 |
2 |
-0.32
|
Bb control |
9.52 |
0 |
0 |
0.5 |
1.25 |
2 |
6.73
|
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