Team:Freiburg/Notebook/12 September
From 2011.igem.org
Contents |
Commons
Minipreps
Investigators: Sandra
Because of the positive testdigest, a miniprep of the PR+GFP was performed. Samples were send for sequencing.
green light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
blue light receptor
Miniprep of PR+lovtap-notgate domain
Investigators: Sandra
Miniprep of:
- 39A
- 39B
- 40A
- 40B
- 41A
- 41B
- 42A
- 42B
- 44A
- 44B
39=PR1, 40=PR2, 41=PR3, 42=PR4, 44=PR6.
2A-assembly: Digestion
Investigators: Sandra
Digestion of the samples from miniprep (10 samples) and digestion of RFP (S1b, S2b).
PR with Lov-Notgate domain:
- 39A
- 39B
- 40A
- 40B
- 41A
- 41B
- 42A
- 42B
- 44A
- 44B
digested with NehI and PstI.
RFP:
- S1b
- S2b
digested with SpeI and PstI.
2A-assembly: Ligation
Ligation of RFP with 39-44.
- 39A-S1b
- 39A-S2b
- 39B-S1b
- 39B-S2b
- 40A-S1b
- 40A-S2b
- 40B-S1b
- 40B-S2b
- 41A-S1b
- 41A-S2b
- 41B-S1b
- 41B-S2b
- 42A-S1b
- 42A-S2b
- 42B-S1b
- 42B-S2b
- 44A-S1b
- 44A-S2b
- 44B-S1b
- 44B-S2b
Trafo
Trafo of the ligated samples on plates with Cm.
red light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
Lysis cassette
High throughput sequencing
Investigators:Theo
All PR parts in C3 vectors, M48 in C3 and also the M48+1 and M48+7 in Amp vectors were sent for sequencing
Precipitator
Testdigest
Name:Rüdiger
| Date: 12.09. |
Continue from Experiment Miniprep (Date) 10.&11.09.
(Name) Rüdiger | |
Project Name: Precipitator |
For one reaction you need: For Mastermix: Number of samples+2extra
4μl | H2O | 280 | |
1μl | Buffer, NEB4 | 70 | |
1μl | BSA (10x) | 70 | |
0,5 μl | Enzym 1 | 35 | Eco R1 |
0,5 μl | Enzym 2 | ||
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!
1aA | 1aB | 1aC | 1bA | 1bB | 1bC | 2aA | 2aB | 2aC |
Eco R1 + Bam H1 | ||||||||
X | X | X | X | X | X | X | X | X |
2bA | 2bB | 2bC | 3bA | 3bB | 3bC | 3aA | 3aB | 3aC |
Eco R1 + Bam H1 | ||||||||
X | X | X | X | X | X | X | X | X |
6aA | 6aB | 6aC | 74aA | 7aB | 7aC | 7bA | 7bB | 7bC |
Eco R1 + Pst 1 | ||||||||
X | X | X | X | X | X | X | X | X |
5bA | 5bB | 5bC | 5aA | 5aB | 5aC | 6bA | 6bB | 6bC |
Eco R1 + Pst 1 | ||||||||
X | X | X | X | X | X | X | X | X |
8a1 | 8a3 | 8a4 | 8b1 | 8b2 | 8b4 | 10a2 | 10a3 | 10a4 |
Eco R1 + Spc 1 | ||||||||
X | X | X | X | X | X | X | X | X |
9a1 | 9a2 | 9a3 | 9a4 | 9b1 | 9b2 | 9b3 | 9b4 |
Eco R1 + Spc 1 | |||||||
X | X | X | X | V | X | X | V |
10b3 | 10b4 | 10b01 | 10b2 | 4a1 | 4a2 | 4a3 | 4b4 |
Eco R1 + Spc 1 | |||||||
V | V | X | V | X | X | V | V |
X=Failure, V=Success
NAME OF YOUR EXPERIMENT
Investigators: NAME