Team:Freiburg/Notebook/18 August
From 2011.igem.org
Contents |
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 Not-Gate, LovTAP in pSB1T3
Investigators: Sophie
Digestion
Amount of DNA and H20:
Sample | DNA μl | H20 μl |
LovTAP | 5 | 33 |
Not-Gate | 2 | 36 |
pSB1T3 | 20 | 18 |
Enzymes necessary for digestion:
LovTAP | Not-Gate | vector | |
enzyme 1 | EcoRI | XbaI | EcoRI |
enzyme 2 | NheI | PstI | PstI |
- Incubation at 37°C for 6 hours
- 20 minutes at 80°C