Team:Paris/Production testing
From 2009.igem.org
(→Part II : Testing delay phase II) |
(→The destabilization of the membrane) |
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|bgcolor="#F8F8F8"|'''PCR :''' | |bgcolor="#F8F8F8"|'''PCR :''' | ||
- | + | pTet: igem plate | |
- | |bgcolor="#E0E3FE"| | + | OmpAs: |
- | |bgcolor="#CBD1FD"| | + | ToRII: pINK2 |
- | |bgcolor="#BCCDFD"| | + | g3p: pINTg3p |
+ | |||
+ | |||
+ | |bgcolor="#E0E3FE"| | ||
+ | |bgcolor="#CBD1FD"|' | ||
+ | |bgcolor="#BCCDFD"| | ||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
Line 558: | Line 563: | ||
ok | ok | ||
- | |bgcolor="#E0E3FE"| | + | |bgcolor="#E0E3FE"|- |
- | + | |bgcolor="#CBD1FD"|- | |
- | - | + | |bgcolor="#BCCDFD"|- |
- | + | ||
- | |bgcolor="#CBD1FD"| | + | |
- | + | ||
- | - | + | |
- | |bgcolor="#BCCDFD"| | + | |
- | |||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
|bgcolor="#F8F8F8"|'''Purification on gel :''' | |bgcolor="#F8F8F8"|'''Purification on gel :''' | ||
- | |||
ok | ok | ||
+ | |bgcolor="#E0E3FE"|- | ||
+ | |bgcolor="#CBD1FD"|- | ||
+ | |bgcolor="#BCCDFD"|- | ||
- | |||
- | |||
- | |bgcolor="# | + | |- style="background: #d8d8d8; text-align: center;" |
+ | |bgcolor="#F8F8F8"|'''Digestion:''' | ||
- | + | pTet S/P | |
- | + | ||
- | + | OmpAs X/P | |
+ | TolRII X/P | ||
- | |||
- | |||
- | + | g3p X/P | |
- | |bgcolor="#E0E3FE"|'''Digestion:''' | + | |
+ | |||
+ | |bgcolor="#E0E3FE"|'''Digestion:''' | ||
+ | pTet-OmpAs S/P | ||
|bgcolor="#CBD1FD"|'''Digestion:''' | |bgcolor="#CBD1FD"|'''Digestion:''' | ||
- | |bgcolor="#BCCDFD"| | + | none |
+ | |||
+ | |bgcolor="#BCCDFD"| | ||
+ | |||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |bgcolor="#F8F8F8"|''' | + | |bgcolor="#F8F8F8"|'''Digestion checking:''' |
ok | ok | ||
- | |bgcolor="#E0E3FE"|''' | + | |bgcolor="#E0E3FE"|'''Digestion checking:''' |
- | + | ok | |
- | + | ||
- | + | ||
- | + | |bgcolor="#CBD1FD"|'''Digestion checking:''' | |
- | |bgcolor="# | + | |
- | - | + | none |
+ | |bgcolor="#BCCDFD"|- | ||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
|bgcolor="#F8F8F8"|'''Ligation:''' | |bgcolor="#F8F8F8"|'''Ligation:''' | ||
- | + | pTet-OmpAs | |
- | + | ||
|bgcolor="#E0E3FE"|'''Ligation:''' | |bgcolor="#E0E3FE"|'''Ligation:''' | ||
+ | none | ||
+ | |||
+ | |||
|bgcolor="#CBD1FD"|'''Ligation:''' | |bgcolor="#CBD1FD"|'''Ligation:''' | ||
+ | none | ||
|bgcolor="#BCCDFD"|'''Ligation:''' | |bgcolor="#BCCDFD"|'''Ligation:''' | ||
Line 629: | Line 634: | ||
|bgcolor="#E0E3FE"|'''Colony PCR:''' | |bgcolor="#E0E3FE"|'''Colony PCR:''' | ||
- | + | ok | |
|bgcolor="#CBD1FD"|'''Colony PCR:''' | |bgcolor="#CBD1FD"|'''Colony PCR:''' | ||
- | + | none | |
|bgcolor="#BCCDFD"|'''Colony PCR:''' | |bgcolor="#BCCDFD"|'''Colony PCR:''' | ||
- | + | none | |
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
|bgcolor="#F8F8F8"|'''Miniprep:''' | |bgcolor="#F8F8F8"|'''Miniprep:''' | ||
- | + | ok | |
|bgcolor="#E0E3FE"|'''Miniprep:''' | |bgcolor="#E0E3FE"|'''Miniprep:''' | ||
- | + | none | |
|bgcolor="#CBD1FD"|'''Miniprep:''' | |bgcolor="#CBD1FD"|'''Miniprep:''' | ||
- | + | none | |
- | |bgcolor="#BCCDFD"| | + | |bgcolor="#BCCDFD"|- |
- | + | ||
- | - | + | |
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
|bgcolor="#F8F8F8"|'''Sequencing :''' | |bgcolor="#F8F8F8"|'''Sequencing :''' | ||
- | + | ok | |
|bgcolor="#E0E3FE"|'''Sequencing :''' | |bgcolor="#E0E3FE"|'''Sequencing :''' | ||
- | + | ok | |
|bgcolor="#CBD1FD"|'''Sequencing :''' | |bgcolor="#CBD1FD"|'''Sequencing :''' | ||
- | + | none | |
|bgcolor="#BCCDFD"|'''Sequencing :''' | |bgcolor="#BCCDFD"|'''Sequencing :''' | ||
- | + | none | |
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" |
Revision as of 02:00, 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 | Ligation: |
Colony PCR :
ok | Colony PCR:
ok | Colony PCR:
none | Colony PCR:
none |
Miniprep:
ok | Miniprep:
none | Miniprep:
none | - |
Sequencing :
ok | Sequencing :
ok | Sequencing :
none | Sequencing :
none |
Stock glycerol:
- | Stock glycerol
- | Stock glycerol
- | Stock glycerol
- |