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Team:Paris/Addressing testing
From 2009.igem.org
iGEM > Paris > WetLab > 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 :
- | Verification on gel :
- |
| Purification on gel :
ok | Purification on gel :
- | Purification on gel :
-
|
| digestion:
ClyA Cterm S/P RFP Nterm X/P PBAD vector : ? E/S
| digestion:
ClyA Cter-RFP Nter vector PSB1A3 E/X | digestion:
- |
| verification digestion:
ok | verification digestion:
ok | verification digestion:
-
|
| ligation:
ClyA Cter-RFP Nter vector PSB1A3 | ligation:
PBAD vector : ? E/S ClyA Cter-RFP Nter vector PSB1A3E/X | ligation:
|
| PCR colo :
ok | PCR colo :
ok | PCR colo :
-
|
| miniprep:
clone : | miniprep:
clone : | miniprep:
-
|
| sequencing :
ok | sequencing :
ok | sequencing :
- |
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
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 |
