04/23/10
- ✓ Minipreps: KAH154-159/MV2
- ✓ Sequencing: KAH156, 157/MV2
Minipreps
> Check with XbaI digests
| Reagent |
Volume
|
Expected:
1. KAH154/MV2 = 1724
2. KAH155/MV2 = 1724
3. KAH156/MV2 = 1724
4. KAH157/MV2 = 1724
5. KAH158/MV2 = 2688
6. KAH159/MV2 = 2691
|
15 μL/lane; 1% agarose
|
| DNA(plasmid) |
2.0 μL
|
| 10X buffer |
1.5
|
| XbaI |
1.0
|
| dH2O |
10.5
|
| |
15 μL --> 37°C/ ~15 min.
|
Sequencing (Genewiz, standard)
--> Can't sequence old KAH156/MV2, used up last of it in transfection
--> Need to confirm 156, 157 in order to have proper id for transfected cells
- KAH156/MV2 (new prep) DDB122: fshPcCD! (should be flyPcCD)
- KAH156/MV2 (new prep) DDB123: fshPcCD! (should be flyPcCD)
- KAH157/MV2 (new prep) DDB122: flyPcCD! (should be fshPcCD)
- KAH157/MV2 (new prep) DDB123: flyPcCD! (should be fshPcCD)
- KAH157/MV2 (old prep) DDB122: bad seq
- KAH157/MV2 (old prep) DDB123: flyPcCD! (should be fshPcCD)
--> Looks like fly and fish PcCD's were mixed up during cloning. Take note of this and continue. May want to check other transfection constructs (including VP64+ set) just to be sure the id's of the stable lines are correct.
| 
Project name
Main project page
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