Team:Freiburg/Notebook/9 August

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


Contents

green light receptor

Results Transformation

We dont see any colonies

  • To-do: do quickchange again.


Quickchange


Investigator:Jakob

PCR


Name:

Jakob

Date:

09.08.2011

Continue from Experiment:
Project Name:

Quickchange CcaR and CcaS

PCR-Mixture for one Reaction:

For a 50 µl reaction use


32,5µl H20
10µl 5x Phusion Buffer
2.5µl Primer fw Primer: P12(CcaR) and P14(CcaS
2.5µl Primer dw Primer: P13(CcaR) and P15(CcaS)
1µl dNTPs
1µl DNA-Template DNA: CcaR and CcaS
0.5 µl Phusion (add in the end)



  • Digest with DpnI


  • Transformation


  • And again, I'm such a fool...

blue light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

Lysis cassette sequencing

Investigators:Theo


Sequencing showed part of the E.coli genome inside the restriction sites where the lysis cassette should be (aaargh)
File:Lys1-P8.gb

Precipitator

new 3A assenbly with Amp Vector

Investigators: Sophie

3A assembly

The last cloning of Theo and Jakob produced one clone with GFP-plastic-binding-domain (GFP-pbd) inside but without Promoter-Ribosome-binding site (PR). So I tried again to combine the GFP-pbd with PRs via 3a assenbly. I used two different PRs (PR4 and PR6) with different RBS binding strenght. Name of the samples: ε5 (GFP-pbd), pSB1A3, PR4, PR6. stored in "Minipreps, verdaut"-box

Ligation

I ligated ε5 (GFP-pbd) to PR4 or PR6 and to the Amp-Vector. Therefore I once took the ratio 3:1 insert to vector and once the ration 1:1 insert to vector.

                       

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