Team:Freiburg/Notebook/3 September

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<a href="https://2011.igem.org/Team:Freiburg/Notebook/2_September">Previous entry</a>
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<a href="https://2011.igem.org/Team:Freiburg/Notebook"> 3 September </a>
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==<span style="color:green;">green light receptor</span>==
==<span style="color:green;">green light receptor</span>==
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<br/>
<br/>
[[File:Freiburg11 09 03Testdigest 2A.Jpg|400px|400px]]
[[File:Freiburg11 09 03Testdigest 2A.Jpg|400px|400px]]
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<br/>
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the samples
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46b= ho1 with a RBS (BBa_B0034)<br/>
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50b= RBS(BBa_B0034)+pcyA+terminator<br/>
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64b= Promotor (BBa_J23110)+CcaS<br/>
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67b= Promotor (BBa_J23110)+CcaR<br/>
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were send on [https://2011.igem.org/Team:Freiburg/Notebook/6_September 6.09] to GATC for sequencing
===3A-assembly:Digestion===
===3A-assembly:Digestion===
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'''InsertI:'''
'''InsertI:'''
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*64
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*64, CcaR
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*46  
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*46, ho1
with EcorI and SpeI
with EcorI and SpeI
'''InsertII:'''
'''InsertII:'''
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*67
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*67, CcaS
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*50
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*50, pcyA
with XbaI and PstI
with XbaI and PstI
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Incubation for 6 hours at 37°C. Vector pSB1K3 was already digested, it only needs to be digested with antarctica for one hour.
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For this assembly we used the constructs from the 2A-assembly made on [https://2011.igem.org/Team:Freiburg/Notebook/31_August 31.08]
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Incubation for 6 hours at 37°C. Vector pSB1C3 was already digested,<br/>  it was just treated with antarctic phosphatase for one hour.
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===3A-assembly:Ligation===
===3A-assembly:Ligation===

Latest revision as of 00:27, 22 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

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.

                       

Retrieved from "http://2011.igem.org/Team:Freiburg/Notebook/3_September"