Team:Freiburg/Notebook/12 September

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Latest revision as of 00:15, 22 September 2011


This is the wiki page
of the Freiburger student
team competing for iGEM 2011.
Thank you for your interest!

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

                       

Retrieved from "http://2011.igem.org/Team:Freiburg/Notebook/12_September"