Team:Paris/Addressing testing

From 2009.igem.org

(Difference between revisions)
(Adressing system)
(Adressing system)
Line 24: Line 24:
-
Time required :  
+
Time required : A lOT !!!!!
Experiments ran :
Experiments ran :
-
PCR : on plasmid ? Oligo TM
+
From plate: mRFP
-
2
+
PCR :
 +
 
 +
ClyA Cterm  plasmid : ? Oligo :  TM :
 +
ClyA Nterm  plasmid : ? Oligo :  TM :
 +
mRFP Cterm  plasmid : ? Oligo :  TM :
 +
mRFP Nterm  plasmid : ? Oligo :  TM :
Verification on gel  
Verification on gel  
 +
ok
 +
Purification on gel  
Purification on gel  
 +
 +
ok
digestion:  
digestion:  
 +
 +
ok
ligation:  
ligation:  
 +
 +
PCR colo :
PCR colo :

Revision as of 11:13, 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 :

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:

ok

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