mm/dd/yyyy
Golden gate 1:10 RhlR and EsaR receivers grew but there were colonies on the backbone plate
Ligation
|
Colonies
|
Bb |
13
|
RhlR |
23
|
EsaR |
14
|
Ran PCR, extension time 3:30 at 50°C
Template
|
F primer
|
R primer
|
pTet-mCh |
130 |
97
|
LuxI GFP rec |
131 |
132
|
LuxR 1:10 colony 1 |
103 |
104
|
LuxR 1:10 colony 2 |
103 |
104
|
LuxR 1:10 colony 3 |
103 |
104
|
LuxR 1:10 colony 4 |
103 |
104
|
LuxR 1:10 colony 5 |
103 |
104
|
LuxR 1:10 colony 5 |
103 |
107
|
Ran above PCR on gel
Template
|
F primer
|
R primer
|
Expected band size
|
Actual band size
|
pTet-mCh |
130 |
97 |
3130 |
3130
|
LuxI GFP rec |
131 |
132 |
610 |
no band
|
LuxR 1:10 colony 1 |
103 |
114 |
1817 |
no band
|
LuxR 1:10 colony 2 |
103 |
114 |
1817 |
1817
|
LuxR 1:10 colony 3 |
103 |
114 |
1817 |
no band
|
LuxR 1:10 colony 4 |
103 |
114 |
1817 |
no band
|
LuxR 1:10 colony 5 |
103 |
114 |
1817 |
1817
|
LuxR 1:10 colony 5 |
103 |
107 |
1035 |
1035
|
picked LuxI and RhlI GFP senders into 4mLs media and shook overnight to screen for successful receiver transformations.
|