Team:Paris/Addressing testing

From 2009.igem.org

(Difference between revisions)
(→Adressing system)
(→Export system)
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[[Image:Protein+Export+Fusion.jpg|650px|center|Production of protein of interest]]
[[Image:Protein+Export+Fusion.jpg|650px|center|Production of protein of interest]]
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Time required : A lOT !!!!!
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Experiments ran :
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{|
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|- style="background: #0a3585; text-align: center; color:white; font-weight:bold; "
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|width= 300px| row 1
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|width=300px| row 2
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|width=300px| row 3
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|- 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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|- style="background: #d8d8d8; text-align: center;"
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|'''Verification on gel :'''
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ok
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|'''Verification on gel :'''
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ok
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|'''Verification on gel :'''
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ok
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|- style="background: #bebebe; text-align: center;" 
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|'''Purification on gel :'''
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ok
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|'''Purification on gel :'''
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ok
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|'''Purification on gel :'''
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ok
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|- style="background: #d8d8d8; text-align: center;"
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|'''digestion:'''
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|'''digestion:'''
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|'''digestion:'''
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|- style="background: #bebebe; text-align: center;" 
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|'''verification digestion:'''
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ok
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|'''verification digestion:'''
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ok
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|'''verification digestion:'''
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ok
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|- style="background: #d8d8d8; text-align: center;"
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|'''ligation:'''
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|'''ligation:'''
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|'''ligation:'''
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|- style="background: #bebebe; text-align: center;" 
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|'''PCR colo :'''
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ok
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|'''PCR colo :'''
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ok
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|'''PCR colo :'''
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ok
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|- style="background: #d8d8d8; text-align: center;"
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|'''miniprep:'''
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clone :
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|'''miniprep:'''
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clone :
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|'''miniprep:'''
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clone :
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|- style="background: #bebebe; text-align: center;" 
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|'''sequencing :'''
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ok
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|'''sequencing :'''
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ok
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|'''sequencing :'''
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ok
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|}

Revision as of 11:45, 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 :

ClyA Cterm plasmid : ? Oligo : TM :

ClyA Nterm plasmid : ? Oligo : TM :

mRFP Cterm plasmid : ? Oligo : TM :

mRFP Nterm plasmid : ? Oligo : TM :


PCR : PCR :


Verification on gel :

ok

Verification on gel :

ok

Verification on gel :

ok

Purification on gel :

ok

Purification on gel :

ok

Purification on gel :

ok


digestion:

ClyA Cterm S/P

RFP Nterm X/P

PBAD E/S


digestion: digestion:
verification digestion:

ok

verification digestion:

ok

verification digestion:

ok


ligation:

ClyA Cter-RFP Nter vector PSB1A3

ligation: ligation:


PCR colo :

ok

PCR colo :

ok

PCR colo :

ok


miniprep:

clone :

miniprep:

clone :

miniprep:

clone :


sequencing :

ok

sequencing :

ok

sequencing :

ok


From plate: mRFP


PCR :



Verification on gel

ok

Purification on gel

ok

digestion:


verification digestion:


ligation:


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


Time required : A lOT !!!!!



Experiments ran :

row 1 row 2 row 3
PCR :



PCR : PCR :


Verification on gel :

ok

Verification on gel :

ok

Verification on gel :

ok

Purification on gel :

ok

Purification on gel :

ok

Purification on gel :

ok


digestion:



digestion: digestion:
verification digestion:

ok

verification digestion:

ok

verification digestion:

ok


ligation:


ligation: ligation:


PCR colo :

ok

PCR colo :

ok

PCR colo :

ok


miniprep:

clone :

miniprep:

clone :

miniprep:

clone :


sequencing :

ok

sequencing :

ok

sequencing :

ok

Retrieved from "http://2009.igem.org/Team:Paris/Addressing_testing"