03/28/15
- Minipreps - BD006/MV2 colonies (from 3/26/15 ligations), 5 mL each (4 total)
Minipreps - BD006/MV2 colonies
- Sigma GenElute kit.
- Elute & back-elute w/ 70 μL elution sln.
- BD006/MV2 plate 1, a
- BD006/MV2 plate 1, b
- BD006/MV2 plate 2, a
- BD006/MV2 plate 2, b
- Restriction Digest check (Fermentas FD)
- Cut with ApaLI (E/S insertion got rid of BB RFC23 cut sites)
Reagent
|
Volume
|
Expected: * BD006/MV2 = 7207, 1243, 288 * MV2 only = 2998, 1243, 288 * BD006/V0120 = 7036, 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! All four look like BD006/MV2
- Submit for sequencing next week
Sample |
OD260 |
260/280 |
ng/μL
|
1. BD006/MV2 1-a |
0.274 |
1.44 |
274.10
|
2. BD006/MV2 1-b |
0.299 |
1.93 |
298.68
|
3. BD006/MV2 2-a |
0.331 |
1.93 |
330.69
|
4. BD006/MV2 2-b |
0.306 |
1.90 |
305.94
|
|