02/10/2014
Ran gel of PCR products from 2/8/14.
Sample
|
Primer 1
|
Primer 2
|
Template
|
Ideal AT
|
Approx Length
|
1 |
P098 |
P099 |
I13522 (2) |
46 |
2372
|
2 |
P098 |
P099 |
I13522 (3) |
46 |
2372
|
3 |
P098 |
P099 |
I13522 9.27 55.7 |
46 |
2372
|
4 |
P103 |
P104 |
I13522 (2) |
49 |
780
|
5 |
P103 |
P104 |
I13522 (3) |
49 |
780
|
6 |
P103 |
P104 |
I13522 9.27 55.7 |
49 |
780
|
- insert gel photo before and after***
Gel extract using Zymo kit. Read concentrations of these and gel extractions from 2/7/14:
Primer 1
|
Primer 2
|
Template
|
260
|
280
|
260/280
|
ng/µL
|
|
|
water |
0.003 |
0.002 |
1.933 |
3.084
|
P103 |
P104 |
I13522 (2) |
0.124 |
0.065 |
1.9 |
124.369
|
P103 |
P104 |
I13522 (3) |
0.123 |
0.065 |
1.9 |
123.324
|
P103 |
P104 |
I13522 9/27/13 55.7 |
0.054 |
0.028 |
1.918 |
54.117
|
P098 |
P099 |
I13522 (3) |
0.098 |
0.051 |
1.931 |
97.574
|
P098 |
P099 |
I13522 (2) |
0.116 |
0.06 |
1.943 |
115.692
|
P117 |
P118 |
K092900 (RhlR) |
0.05 |
0.027 |
1.874 |
50.159
|
P105 |
P106 |
R0011 (2) |
0.005 |
0.002 |
2.632 |
5.229
|
P113 |
P114 |
LuxR (F2620) |
0.057 |
0.029 |
1.968 |
57.356
|
P105 |
P106 |
R0011 (1) |
0.002 |
0 |
-6.667 |
2.073
|
P088 |
P102 |
Receiver vector |
0.113 |
0.06 |
1.896 |
113.028
|
|
Golden gate to build LuxR receiver
Part
|
Length (bp)
|
Concentration (ng/ul)
|
fmol/ul
|
Final volume to dilute 1ul to get 20 fmol/ul
|
Receiver vector |
2372 |
113 |
72.2 |
3.6
|
Lux box |
56 |
20 |
541.1 |
27.1
|
-35-B0033-GFP-TT |
945 |
124 |
198.8 |
9.9
|
R0011-RBS |
85 |
5 |
89.1 |
4.5
|
LuxR |
817 |
57 |
105.7 |
5.3
|
GG to build RhlR receiver
Part
|
Length (bp)
|
Concentration (ng/ul)
|
fmol/ul
|
'
|
Receiver vector |
2372 |
113 |
72.2 |
3.6
|
Rhl box |
56 |
20 |
541.1 |
27.1
|
-35-B0033-GFP-TT |
945 |
124 |
198.8 |
9.9
|
R0011-RBS |
85 |
5 |
89.1 |
4.5
|
RhlR |
765 |
50 |
99.0 |
5.0
|
|