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Team:Paris/Production testing
From 2009.igem.org
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Revision as of 02:08, 22 October 2009
iGEM > Paris > WetLab > Production
The delay
We separate the construction on two plasmids : Part I and Part II.
Part I : Testing delay phase I
Experiments ran :
| Column 1 | Column 2 | Column 3 | Column 4 |
| PCR :
pBAD: plate igem
| - | - | - |
| Verification on gel :
ok | - | - | - |
| Purification on gel :
ok | - | - | -
|
| Digestion:
pBAD S/P
| Digestion:
pBAD-LacI E/S
| Digestion:
RBS-YFP-ter E/X | - |
| Verification digestion:
ok | Verification digestion:
ok | Verification digestion:
ok | -
|
| Ligation:
pBAD-LacI RBS-YFP | Ligation: | Ligation: | Ligation: |
| Colony PCR :
ok | Colony PCR :
ok | Colony PCR :
ok | Colony PCR:
ok |
| Miniprep:
clone | Miniprep:
clone | Miniprep:
clone | Miniprep:
clone |
| Sequencing :
ok | Sequencing :
ok | Sequencing :
ok | Sequencing :
ok |
| Stock glycerol:
| Stock glycerol
Cly A(Cter)-(Nter)RFP: S72(clone 8) | Stock glycerol
- | Stock glycerol
- |
Part I : Functional testing of the part I
PBAD LacI LVA- YFP was transforme into Top ten Bacteria and grew on medium :
1% arabinose
1% glucose
LB pure
Part II : Testing delay phase II
Experiments ran :
| Column 1 | Column 2 | Column 3 | Column 4 |
| PCR :
terminator: igem plate pLac: igem plate
| - | - | - |
| Verification on gel :
ok | -
- | -
- | -
- |
| Purification on gel :
ok |
- |
- |
-
|
| Digestion:
pLac S/P
| Digestion:
plac-TetR E/S pTet-mRFP X/P | Digestion:
pLac-tetR-ter S/P
| Digestion:
pLac-tetR-ter-pTet-mRFP E/P
|
| Digestion checking:
ok | Digestion checking:
ok | Digestion checking:
ok | Digestion checking:
ok
|
| Ligation:
pLac-tetR
| Ligation:
pLac-TetR-ter | Ligation:
pLac-ter-pTet-mRFP | Ligation:
pLac-ter-pTet-mRFP-pSB3T5
|
| Colony PCR:
ok | Colony PCR :
ok | Colony PCR:
ok | Colony PCR :
ok |
| Miniprep:
ok | Miniprep:
ok | Miniprep:
ok | Miniprep:
ok |
| Sequencing :
ok | Sequencing :
ok | Sequencing :
ok | Sequencing :
ok |
| Stock glycerol:
- | Stock glycerol
- | Stock glycerol
- | Stock glycerol
- |
Part II : Functional testing of the part II
Ptet RFP Plac TetR was tranforme into Top10 bacteria and grew on media :
0,1% IPTG .
1% glucose.
LB pure.
The destabilization of the membrane
Experiments ran :
| Column 1 | Column 2 | Column 3 | Column 4 |
| PCR :
pTet: igem plate OmpAs: ToRII: pINK2 g3p: pINTg3p
| ' | ||
| Verification on gel :
ok | - | - | -
|
| Purification on gel :
ok | - | - | -
|
| Digestion:
pTet S/P OmpAs X/P
| Digestion:
pTet-OmpAs S/P | Digestion:
none | -
|
| Digestion checking:
ok | Digestion checking:
ok | Digestion checking:
none | -
|
| Ligation:
pTet-OmpAs | Ligation:
none
| Ligation:
none | - |
| Colony PCR :
ok | Colony PCR:
none | Colony PCR:
none | - |
| Miniprep:
ok | Miniprep:
none | Miniprep:
none | - |
| Sequencing :
ok | Sequencing :
none | Sequencing :
none | -
|
| Stock glycerol:
- | Stock glycerol
none | Stock glycerol
none | - |
