09/18/14 - Meli'sa Crawford
- Plasmid sequencing - planning
Plasmid sequencing - planning
- P0001 (blue cap = 100 μM)
- P0002 (blue cap = 100 μM)
- Thaw stock primers (blue capped tubes) at room temp.
- Get two sterile 1.5 mL tubes
- Label each with the primer name and "10 μM"
- Add 90 μL dH2O to each tube
- Transfer 10 μL P0001 from blue caped tube into 90 μL H2O in new tube. Final volume = 100 μL, final conce. = 10 μM
- Repeat last step for P0002.
Tube |
Primer (1 μL of 10 μM) |
DNA (200 ng) |
[DNA] |
Vol DNA |
Vol 10 μM primer |
Vol dH2O
|
1. |
P0001 |
BD003 |
128 ng/μL |
1.6 μL |
1.0 μL |
7.4 μL
|
2. |
P0002 |
BD003 |
128 ng/μL |
1.6 μL |
1.0 μL |
7.4 μL
|
3. |
P0001 |
BD004 |
141 ng/μL |
1.4 μL |
1.0 μL |
7.6 μL
|
4. |
P0002 |
BD004 |
141 ng/μL |
1.4 μL |
1.0 μL |
7.6 μL
|
5. |
P0001 |
BD005 |
290.2 ng/μL |
0.7 μL |
1.0 μL |
8.3 μL
|
6. |
P0002 |
BD005 |
290.2 ng/μL |
0.7 μL |
1.0 μL |
8.3 μL
|
7. |
P0001 |
BD006 |
255.3 ng/μL |
0.8 μL |
1.0 μL |
8.2 μL
|
8. |
P0002 |
BD006 |
255.3 ng/μL |
0.8 μL |
1.0 μL |
8.2 μL
|
- Use 0.2 mL PCR strip tubes
- Be sure to leabel each one
- Place in a plastic baggie (recycled)
- Use tape to secure baggie with tubes to order form
|