Team:Freiburg/Notebook/3 September

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of the Freiburger student
team competing for iGEM 2011.
Thank you for your interest!

Contents

green light receptor

Testdigestion of 2a-assembly

Investigators: Julia
Continuing from 31 of Aug. [1]
Before Miniprep of 20 different colonies.
Each minipreped DNA-sample (3μl)was digested with EcoRI and PstI.

Result:

Freiburg11 09 03Testdigest 2A.Jpg


the samples

46b= ho1 with a RBS (BBa_B0034)

50b= RBS(BBa_B0034)+pcyA+terminator

64b= Promotor (BBa_J23110)+CcaS

67b= Promotor (BBa_J23110)+CcaR

were send on 6.09 to GATC for sequencing

3A-assembly:Digestion

Investigators: Sandra

Digestion of:

InsertI:

  • 64, CcaR
  • 46, ho1

with EcorI and SpeI

InsertII:

  • 67, CcaS
  • 50, pcyA

with XbaI and PstI

For this assembly we used the constructs from the 2A-assembly made on 31.08

Incubation for 6 hours at 37°C. Vector pSB1C3 was already digested,
it was just treated with antarctic phosphatase for one hour.

3A-assembly:Ligation

Investigators: Julia

blue light receptor

Stop of ligation

Investigators: Sandra

Inactivation of ligase at 80°C for 20 minutes.
Stored in the blue light box, labelled: ♥C3 Lig., and Not C3 Lig.

To do: Trafo


Transformation

with ♥C3 Lig. and Not C3 Lig

Investigator: Sophie

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME


Precipitator

Picking clones

Investigators: Sandra

Only few colonies grew on two plates. These clones were picked and incubated in LB with amp at 37°C.

                       

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