08/22/11
- ✓ Cultures/minipreps: KAH235/V0200 (2)
- ✓ Midipreps: Pc-TF/pSB31 constructs for Manching
Minipreps
> Check with N/S digests
| Reagent |
Volume
|
Expected:
1,2. KAH235/V0200 = 4442, 2787
|
Miniprep digests
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.
|
Midipreps
> Use QIAGEN Midiprep Plus kit
> Check with NcoI digests
| Reagent |
Volume
|
Expected:
1. KAH235/V0200 = size
|
Midiprep digests
2 μL DNA, 15 μL/lane; 1% agarose
5 μL DNA, 15 μL/lane; 1% agarose
|
| DNA(plasmid) |
2.0 / 5.0 μL
|
| 10X buffer |
1.5
|
| NcoI |
1.0
|
| dH2O |
10.5 / 7.5
|
| |
15 μL --> 37°C/ ~15 min.
|
--> Nanodrop reading showed 150 - 270 ng/μL for these preps, but no DNA is visible on the gel. Try re-transformation, followed by midi prep.
|
Project name
Main project page
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