04/10/15
- Minipreps - BD004/MV2 (6x 5mL cultures)
Minipreps - BD004/MV2
- ---Karmella 20:52, 10 April 2015 (EDT):
- Sigma GenElute kit.
- Elute & back-elute w/ 70 μL elution sln.
- BD004/MV2 a
- BD004/MV2 b
- BD004/MV2 c
- BD004/MV2 d
- BD004/MV2 e
- BD004/MV2 f
- Restriction Digest check (Fermentas FD)
- Cut with ApaLI (E/S insertion got rid of BB RFC23 cut sites)
Reagent
|
Volume
|
Expected: * BD004/MV2 = 7108, 1243, 288 * MV2 only = 2998, 1243, 288 * BD004/V0120 = 5659, 1246, 497
|
30 μL/lane, 1% agarose; Ladder
|
DNA (plasmid) |
3.0 μL
|
10x buffer |
1.5
|
ApaLI |
1.0
|
dH2O |
9.5
|
|
15 μL --> 37°C/ ~10 min.
|
- Conclusions
- Success, and very lucky! Clone "a" is the desired plasmid.
- Keep clone a and discard other minipreps.
- Mark the streak library accordingly.
Sample |
OD260 |
260/280 |
ng/μL
|
1. BD004/MV2 a |
0.400 |
1.90 |
399.66
|
2. BD004/MV2 b |
0.344 |
1.90 |
344.33
|
3. BD004/MV2 c |
0.271 |
1.88 |
270.79
|
4. BD004/MV2 d |
0.292 |
1.89 |
292.47
|
5. BD004/MV2 e |
0.279 |
1.90 |
279.28
|
6. BD004/MV2 f |
0.221 |
1.88 |
220.53
|
|