03/19/10
- ✓ Minipreps: KAH142, 96/V0201 (2 each)
- ✓ Sequencing order: KAH96/V0201
- ✓ Cultures: KAH142/V0201 (8)
Minipreps
> Check with XbaI digest
> XbaI fragment will have an extra 641 bp from vector sequence; remaining vector = 4516
| Reagent |
Volume
|
Expected:
1. KAH142/V0201 = 4571 (5212)
2. KAH96/V0201 = 2825 (3466)
|
15 μL/lane; 1% agarose
|
| DNA(plasmid) |
2.0 μL
|
| 10X buffer |
1.5
|
| EcoRI |
1.0
|
| dH2O |
10.5
|
| |
15 μL --> 37°C/ ~15 min.
|
--> Sequence KAH96/V0201 plasmids
--> Pick 8 new colonies for KAH146/V0201
Sequencing order
- KAH96_V0201-1 3T3: HSVtkTATA-Kozak is missing
- KAH96_V0201-1 BGHR: good
- KAH96_V0201-2 3T3: HSVtkTATA-Kozak is missing
- KAH96_V0201-2 BGHR: good
--> Minimal promoter is missing! KAH142 might also be bad. Sequence construct intermediates KAH86 and 96 in V0120.
| 
Project name
Main project page
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