Team:Paris/Addressing testing

From 2009.igem.org

(Difference between revisions)
(Adressing system)
(Adressing system)
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|- style="background: #bebebe; text-align: center;"
|- style="background: #bebebe; text-align: center;"
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|PCR  
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|PCR :
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|PCR
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|PCR :
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|PCR
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|PCR :
|- style="background: #d8d8d8; text-align: center;"
|- style="background: #d8d8d8; text-align: center;"
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|height=40px|[]
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|Verification on gel :
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|2003
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|Verification on gel :
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|Miller SI & Guina T.
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|Verification on gel :
|- style="background: #bebebe; text-align: center;"   
|- style="background: #bebebe; text-align: center;"   
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|height=40px|[]
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|Purification on gel :
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|2005
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|Purification on gel :
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|Kuehn MJ & Kesty NC.
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|Purification on gel :
|- style="background: #d8d8d8; text-align: center;"
|- style="background: #d8d8d8; text-align: center;"
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|height=40px|[]
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|digestion:
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|2006
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|digestion:
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|McBroom AJ & Kuehn MJ.
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|digestion:
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|verification digestion:
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|2007
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|verification digestion:
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|McBroom AJ & Kuehn MJ.
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|verification digestion:
|- style="background: #d8d8d8; text-align: center;"
|- style="background: #d8d8d8; text-align: center;"
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|height=40px|[]
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|ligation:
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|2009
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|ligation:
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|Deatherage BL & Cookson BT.
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|ligation:
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|PCR colo :
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|2009
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|PCR colo :
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|Purnick PE & Weiss R.
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|PCR colo :
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|- style="background: #d8d8d8; text-align: center;"
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|miniprep:
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|miniprep:
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|miniprep:
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|sequencing :
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|sequencing :
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PBAD E/S
PBAD E/S
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verification digestion:

Revision as of 11:34, 17 October 2009

iGEM > Paris > WetLab > Addressing

WetLab

Main - Addressing - Production - reception -
Addressing


Adressing system

global constructions :


plasmid = PSB3T5


Time required : A lOT !!!!!



Experiments ran :

row 1 row 2 row 3
PCR : PCR : PCR :


Verification on gel : Verification on gel : Verification on gel :


Purification on gel : Purification on gel : Purification on gel :
digestion: digestion: digestion:
verification digestion: verification digestion: verification digestion:


ligation: ligation: ligation:


PCR colo : PCR colo : PCR colo :
miniprep: miniprep: miniprep:


sequencing : sequencing : sequencing :


From plate: mRFP


PCR :


ClyA Cterm plasmid : ? Oligo : TM : ClyA Nterm plasmid : ? Oligo : TM : mRFP Cterm plasmid : ? Oligo : TM : mRFP Nterm plasmid : ? Oligo : TM :


Verification on gel

ok

Purification on gel

ok

digestion:

ClyA Cterm S/P RFP Nterm X/P PBAD E/S

verification digestion:


ligation:

ClyA Cter-RFP Nter vector PSB1A3

PCR colo :

miniprep:

verifivation of the miniprep by digestion

sequencing [[ClyA]]

-ClyA Nterm= biobrick+sequencage+stock glycerol Done

-ClyA Cterm= biobrick+sequencage+stock glycerol Done

[[-RFP-]]

-RFP Nterm= biobrick+sequencage+stock glycerol Done

-RFP Cterm= biobrick+sequencage+stock glycerol Done

[[-ClyA Cter-RFP Nter-]]

-ClyA Cter-RFP Nter= biobrick+stock glycerol Done

[[-RFP Cter-ClyA Nter-]]

-miss ligation between 2 parts (j'ai fait plusieurs ligation le 7ou8/09, mais pas marché le 9/09 apres culture overnight, voir si les autres ont continué)


[[PBad-ClyA Cter-RFP Nter-]]

-failed many time with:

Pbad cut in S/P and ClyA Cter-RFP Nter in X/P

Pbad cut in E/S and ClyA Cter-RFP Nter in E/X


Export system

Production of protein of interest



Production of protein of interest