Team:Paris/Production testing
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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 lacI E/S terminator E/X RBS S/P YFP X/P | Digestion:
pBAD-LacI E/S RBS-YFP E/S | Digestion:
RBS-YFP-ter E/X | Digestion: |
Verification digestion:
ok | Verification digestion:
ok | Verification digestion:
ok | -
|
- | Ligation: | Ligation: | - |
- | Colony PCR :
ok | Colony PCR :
- | Colony PCR:
- |
Miniprep:
STOPPED | Miniprep:
- | Miniprep:
- | Miniprep:
- |
Sequencing :
- | Sequencing :
- | Sequencing :
- | Sequencing :
- |
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 :
mRFP Cterm matrix : pSB1A3 Oligo :O59 and O60 TM :
| PCR : | PCR : | PCR : |
Verification on gel :
ok | Verification on gel :
- | Verification on gel :
- | Verification on gel :
- |
Purification on gel :
ok | Purification on gel :
- | Purification on gel :
- | Purification on gel :
-
|
Digestion:
TatABCE | Digestion: | Digestion: | Digestion: |
Verification digestion:
ok | Verification digestion:
- | Verification digestion:
- | Verification digestion:
-
|
Ligation:
TatABCE | Ligation: | Ligation: | Ligation: |
Colony PCR:
ok | Colony PCR :
- | Colony PCR:
- | Colony PCR :
- |
Miniprep:
STOPPED | Miniprep:
- | Miniprep:
- | Miniprep:
- |
Sequencing :
- | Sequencing :
- | Sequencing :
- | Sequencing :
- |
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 :
TatABCE matrix : | PCR : | PCR : | PCR : |
Verification on gel :
ok | Verification on gel :
- | Verification on gel :
- | Verification on gel :
- |
Purification on gel :
ok | Purification on gel :
- | Purification on gel :
- | Purification on gel :
-
|
Digestion:
TatABCE | Digestion: | Digestion: | Digestion: |
Verification digestion:
ok | Verification digestion:
- | Verification digestion:
- | Verification digestion:
-
|
Ligation:
TatABCE | Ligation: | Ligation: | Ligation: |
Colony PCR :
ok | Colony PCR:
- | Colony PCR:
- | Colony PCR:
- |
Miniprep:
STOPPED | Miniprep:
- | Miniprep:
- | Miniprep:
- |
Sequencing :
- | Sequencing :
- | Sequencing :
- | Sequencing :
- |
Stock glycerol:
- | Stock glycerol
- | Stock glycerol
- | Stock glycerol
- |