Team:Freiburg/Notebook/18 August

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*Incubation at 37°C for 6 hours
*Incubation at 37°C for 6 hours
*20 minutes at 80°C
*20 minutes at 80°C
 +
<br/>
 +
'''I won't continue with the IPTG-inducible-promoter-experiments because the part is shit.'''
 +
We will work with a new IPTG-promoter
{{:Team:Freiburg/Templates/footer}}
{{:Team:Freiburg/Templates/footer}}

Revision as of 14:19, 18 August 2011


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

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 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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