green light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
blue light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
red light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
Lysis cassette
NAME OF YOUR EXPERIMENT
Investigators:NAME
Precipitator
Miniprep
zymo Kit
| Name:Sophie
| Date:18.08.11
|
| Continue from Experiment: Cloning: ε5 was empty and the products from the cloning of 12.08.11 contain GFP-pbd too. (Date): 12.08.11
(Name): Sophie
|
| Project Name: inducible Promoter with pbd with cm-vector
|
Documentation:
Why are you doing this experiment? Name the parts which you extract.
ε5 was empty and the products from the cloning of 12.08.11 contain GFP-pbd too.
Name of the parts: GFP-pbd 4 4 4, GFP-pbd 4 1 3, GFP-pbd 6 6 8, GFP-pbd 4 2 2
Describe your results and mistakes and measure the DNA concentration with the Nanodrop and note the results.
| GFP-pbd 4 4 4: 164,1 ng/µl
GFP-pbd 4 1 3: 100,8 ng/µl
GFP-pbd 6 6 8: 163,5 ng/µl
GFP-pbd 4 2 2: 144,4 ng/µl
|
How did you label your probes and where are they stored?
| Labelled GFP-pbd 4 4 4, GFP-pbd 4 1 3, GFP-pbd 6 6 8, GFP-pbd 4 2 2
stored in -20 until sequencing
|
Testdigest
| Name: Sophie
| Date: 18.08.11
|
| Continue from Experiment: Miniprep (Date): 18.08.11
(Name): Sophie
|
| Project Name:inducible promoter for pbd with cm-vector
|
For one reaction you need: For Mastermix: Number of samples+2extra
| 4μl
| H2O
| 20
|
|
| 1μl
| Buffer, NEB4
| 5
|
|
| 1μl
| BSA (10x)
| 5
|
|
| 0,5 μl
| Enzym 1
| 2,5
|
|
| 0,5 μl
| Enzym 2
| 2,5
|
|
| 3 μl
| DNA
|
|
|
10 μl total volume
Give 3 μl of DNA in an eppi and add 7μl of the mastermix.
Incubate for about 1h at 37°C.
Add 1 μl Loading dye buffer and load the gel.
Take a picture of the gel, print picture and label the lanes!
3A-assembly of IPTG-promoter, pbd-GFP and Cm-vector
Investigators: Sophie
Digestion
Amount of DNA and H20:
| Sample
| DNA μl
| H20 μl
|
| IPTG-Promoter
| 2,5
| 33
|
| GFP-pbd
| 3
| 36
|
| pSB1A3
| 20
| 18
|
Enzymes necessary for digestion:
|
| GFP-pbd: 4 4 4 or 6 6 8
| IPTG-Promoter: S54
| vector: psB1C3
|
| enzyme 1
| EcoRI
| XbaI
| EcoRI
|
| enzyme 2
| NheI
| PstI
| PstI
|
- Incubation at 37°C for 6 hours
- 20 minutes at 80°C
I won't continue with the IPTG-inducible-promoter-experiments because the part is shit.
We will work with a new IPTG-promoter
"
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