05/30/11
- ✓ Miniprep digests: check miR sensor set (re-transformed)
Minipreps
> Check with E/P digests
| Reagent |
Volume
|
Expected:
MV2 = 4529
1,2. FC1 = 8063
3,4. FC2 = 8255
5,6. FC3 = 7991
7. FC4 = 8183
8. FC31 = 3052
9. FC32 = 2980
10. FC33 = 3244
11. FC34 = 3172
12,13. KAH118 = 7930
14,15. KAH119 = 8002
16. KAH100 = 3085
17. KAH101 = 3157
18,19. KAH121 = 8102
20,21. KAH122 = 8174
22. KAH124 = 8174
23,24. KAH125 = 8246
|
15 μL/lane; 1% agarose
15 μL/lane; 1% agarose
|
| DNA(plasmid) |
2.0 μL
|
| 10X buffer |
1.5
|
| EcoRI |
1.0
|
| PstI |
1.0
|
| dH2O |
9.5
|
| |
15 μL --> 37°C/ ~15 min.
|
-> Note: all worked except for FC1, KAH118, KAH119, KAH124. Use retransformations (#1, 2, 12, 13, 14, 15, 22) to grow 5mL cultures and try again.
--> Result: Miniprep digests do not look good for FC1, KAH118, KAH119, but spec readings show ~160 ng/μL. Try one of each for transfection to see what happens.
-> Spec readings show good concentrations for all others. Keep for U2OS transfectons.
-> Made streak plate from 2 mL cultures yesterday.
|
Project name
Main project page
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