Team:Paris/Addressing testing

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Revision as of 11:44, 17 October 2009

iGEM > Paris > WetLab > Addressing

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WetLab

Main - Addressing - Production - reception - Addressing

Adressing system

global constructions :

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

@@ Line 81: / Line 81: @@
 |- style="background: #d8d8d8; text-align: center;"
-|digestion:
+|'''digestion:'''
-|digestion:
-|digestion:
+ClyA Cterm S/P
+RFP  Nterm X/P
+PBAD E/S
+|'''digestion:'''
+|'''digestion:'''
 |- style="background: #bebebe; text-align: center;"
-|verification digestion:
+|'''verification digestion:'''
-|verification digestion:
-|verification digestion:
+ok
+|'''verification digestion:'''
+ok
+|'''verification digestion:'''
+ok
 |- style="background: #d8d8d8; text-align: center;"
-|ligation:
+|'''ligation:'''
-|ligation:
-|ligation:
+ClyA Cter-RFP Nter  vector PSB1A3
+|'''ligation:'''
+|'''ligation:'''
 |- style="background: #bebebe; text-align: center;"
-|PCR colo :
+|'''PCR colo :'''
-|PCR colo :
-|PCR colo :
+ok
+|'''PCR colo :'''
+ok
+|'''PCR colo :'''
+ok
 |- style="background: #d8d8d8; text-align: center;"
-|miniprep:
+|'''miniprep:'''
-|miniprep:
-|miniprep:
+clone :
+|'''miniprep:'''
+clone :
+|'''miniprep:'''
+clone :
 |- style="background: #bebebe; text-align: center;"
-|sequencing :
+|'''sequencing :'''
-|sequencing :
-|sequencing :
+ok
+|'''sequencing :'''
+ok
+|'''sequencing :'''
+ok
 |}
@@ Line 134: / Line 179: @@
 digestion:
-ClyA Cterm S/P
-RFP  Nterm X/P
-PBAD E/S
 verification digestion:
@@ Line 143: / Line 185: @@
 ligation:
-ClyA Cter-RFP Nter  vector PSB1A3
 PCR colo :