Team:Stony Brook/Notebook/Cancer-W11
Week 11: 9/5 - 9/11
Digest of CR-1 Plasmids
| Content |
Experimental ul |
Control ul |
| H2O |
15.5 |
16.5 |
| Cutsmart |
2.5 |
2.5 |
| XhoI |
0.5 |
X |
| EcoRI-HF |
0.5 |
X |
| DNA |
6 |
6 |
PCR of Toggle 2
PCR
| Contents |
Trial 1 (ul) |
Trial 2 (ul) |
Q5 MasterMix |
12.5 |
12.5 |
| H2O |
6.5 |
5.5 |
| 10 uM F Primer |
0.5 |
1.0 |
| 10 uM R Primer |
0.5 |
1.0 |
| 100pg/ul Construct |
5 |
5 |
PCR Settings: 30 cycles
| Phase |
Temperature (°C) |
Time (sec) |
| Initial Denaturation |
98 |
30 |
| Denaturation |
98 |
10 |
| Annealing |
68 |
20 |
| Extension |
72 |
60 |
| Final Extension |
72 |
120 |
Ran Digests
Digest of J6100
| Content |
Experimental ul |
| H2O |
7.5 |
| Cutsmart |
1.0 |
| XbaI |
0.5 |
| DNA |
1.0 |
Digest of CR-1
| Content |
Experimental ul |
Control ul |
| H2O |
5.0 |
6.0 |
| Cutsmart |
1.0 |
1.0 |
| EcoRI-HF |
0.5 |
X |
| XbaI |
0.5 |
X |
| DNA |
3 |
3 |
Digest of Dean Vector #3
| Content |
Experimental ul |
Control ul |
| H2O |
2.0 |
3.0 |
| Cutsmart |
1.0 |
1.0 |
| EcoRI-HF |
0.5 |
X |
| XhoI |
0.5 |
X |
| Vector |
6 |
6 |
PCR
| Contents |
Trial 1 (ul) |
Trial 2 (ul) |
Phire |
25.0 |
25.0 |
25.0 |
| H2O |
21.5 |
19.0 |
14.0 |
| 10 uM F Primer |
1.25 |
1.50 |
5.0 |
| 10 uM R Primer |
1.25 |
1.50 |
5.0
|
| 100pg/ul Construct |
1 |
1 |
1 |
PCR Settings: 30 cycles
| Phase |
Temperature (°C) |
Time (sec) |
| Initial Denaturation |
98 |
30 |
| Denaturation |
98 |
5 |
| Annealing |
66 |
10 |
| Extension |
72 |
20 |
| Final Extension |
72 |
60 |
