Team:Freiburg/Notebook/11 August

From 2011.igem.org

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===Tramsformation with inducible Promoter===
===Tramsformation with inducible Promoter===
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===='''Investigators: Sophie'''====
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'''Investigators: Sophie'''
We didn't get any clones with both Plastic binding domain and Promoter-RBS inside, the Pbd seems to be toxic when expressed.So we want to try if it is possible to get cells with an inducible Promoter and Pbd. In this case we could cultivate the cells and induce expression of the Pbd only shortly before lysis. As our Light-inducible Promoters are not finished we use the IPTG inducible Promoter BBa_J04500 (Plate4, 12A)
We didn't get any clones with both Plastic binding domain and Promoter-RBS inside, the Pbd seems to be toxic when expressed.So we want to try if it is possible to get cells with an inducible Promoter and Pbd. In this case we could cultivate the cells and induce expression of the Pbd only shortly before lysis. As our Light-inducible Promoters are not finished we use the IPTG inducible Promoter BBa_J04500 (Plate4, 12A)
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 +
===PCR===
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 +
'''Investigators: Rüdiger'''
 +
 +
PCR of precipitator.
 +
 +
Primer fw:
 +
*p51
 +
*p52
 +
*p53
 +
 +
Primer dw:
 +
*p54
 +
 +
DNA template: S43
 +
 +
DMSO was used to reduce Tm about 5°C.
 +
 +
 +
{{:Team:Freiburg/Templates/footer}}
{{:Team:Freiburg/Templates/footer}}

Revision as of 13:01, 13 September 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

PCR

Name: Sophie


Date: 11.8.11
Continue from Experiment: new experiment


Project Name: changes in the LovTAP for 3A assembly

PCR-Mixture for one Reaction:

For a 50 µl reaction use


32,5µl H20 Name
10µl 5x Phusion Buffer of Primer
2.5µl Primer fw
2.5µl Primer dw
1µl dNTPs of Template DNA
1µl DNA-Template
0.5 µl Phusion (add in the end)

What program do you use? Normal PCR with 60°C annealing temperature


To confirm the PCR-Product has the correct size, load 2 µl of the sample onto an agarose-gel.


How did you label the PCR-Product, where is it stored and what do you do next?

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME


Precipitator

Tramsformation with inducible Promoter

Investigators: Sophie We didn't get any clones with both Plastic binding domain and Promoter-RBS inside, the Pbd seems to be toxic when expressed.So we want to try if it is possible to get cells with an inducible Promoter and Pbd. In this case we could cultivate the cells and induce expression of the Pbd only shortly before lysis. As our Light-inducible Promoters are not finished we use the IPTG inducible Promoter BBa_J04500 (Plate4, 12A)

PCR

Investigators: Rüdiger

PCR of precipitator.

Primer fw:

  • p51
  • p52
  • p53

Primer dw:

  • p54

DNA template: S43

DMSO was used to reduce Tm about 5°C.


                       

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