Team:Paris/Production testing
From 2009.igem.org
(→Part I : Testing delay phase I) |
(→Part II : Functional testing of the part II) |
||
(8 intermediate revisions not shown) | |||
Line 237: | Line 237: | ||
|bgcolor="#CBD1FD"|'''Sequencing :''' | |bgcolor="#CBD1FD"|'''Sequencing :''' | ||
- | + | ok | |
|bgcolor="#BCCDFD"|'''Sequencing :''' | |bgcolor="#BCCDFD"|'''Sequencing :''' | ||
- | + | ok | |
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
|bgcolor="#F8F8F8"| '''Stock glycerol:''' | |bgcolor="#F8F8F8"| '''Stock glycerol:''' | ||
Line 267: | Line 267: | ||
LB pure | LB pure | ||
+ | |||
+ | |||
+ | To learn more, see the [[Team:Paris/Conclusion | Conclusion page]] | ||
+ | |||
+ | |||
Line 307: | Line 312: | ||
|bgcolor="#F8F8F8"|'''PCR :''' | |bgcolor="#F8F8F8"|'''PCR :''' | ||
- | + | terminator: igem plate | |
+ | pLac: igem plate | ||
- | |||
- | |bgcolor="#E0E3FE"| | + | TetR |
- | |bgcolor="#CBD1FD"| | + | |
- | |bgcolor="#BCCDFD"| | + | |
+ | pTet: | ||
+ | |||
+ | |||
+ | mRFP | ||
+ | |||
+ | |||
+ | |bgcolor="#E0E3FE"|- | ||
+ | |bgcolor="#CBD1FD"|- | ||
+ | |bgcolor="#BCCDFD"|- | ||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
Line 321: | Line 335: | ||
ok | ok | ||
- | |bgcolor="#E0E3FE"| | + | |bgcolor="#E0E3FE"|- |
- | - | ||
- | |bgcolor="#CBD1FD"| | + | |bgcolor="#CBD1FD"|- |
- | - | ||
- | |bgcolor="#BCCDFD"| | + | |bgcolor="#BCCDFD"|- |
- | - | ||
Line 337: | Line 351: | ||
ok | ok | ||
- | |bgcolor="#E0E3FE"| | + | |bgcolor="#E0E3FE"| |
- | - | ||
- | |bgcolor="#CBD1FD"| | + | |bgcolor="#CBD1FD"| |
- | - | ||
- | |bgcolor="#BCCDFD"| | + | |bgcolor="#BCCDFD"| |
- | - | ||
Line 353: | Line 367: | ||
|bgcolor="#F8F8F8"|'''Digestion:''' | |bgcolor="#F8F8F8"|'''Digestion:''' | ||
- | + | pLac S/P | |
+ | |||
+ | |||
+ | TetR X/P | ||
+ | |||
+ | |||
+ | terminator E/X | ||
+ | |||
+ | |||
+ | pTet S/P | ||
+ | |||
+ | |||
+ | mRFP X/P | ||
+ | |||
+ | |||
+ | pSB3T5 E/P | ||
|bgcolor="#E0E3FE"|'''Digestion:''' | |bgcolor="#E0E3FE"|'''Digestion:''' | ||
+ | plac-TetR E/S | ||
+ | |||
+ | pTet-mRFP X/P | ||
+ | |||
|bgcolor="#CBD1FD"|'''Digestion:''' | |bgcolor="#CBD1FD"|'''Digestion:''' | ||
+ | pLac-tetR-ter S/P | ||
+ | |||
+ | |||
|bgcolor="#BCCDFD"|'''Digestion:''' | |bgcolor="#BCCDFD"|'''Digestion:''' | ||
+ | pLac-tetR-ter-pTet-mRFP E/P | ||
+ | |||
|- 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"|''' | + | |bgcolor="#CBD1FD"|'''Digestion checking:''' |
- | + | ok | |
- | |bgcolor="#BCCDFD"|''' | + | |bgcolor="#BCCDFD"|'''Digestion checking:''' |
- | + | ok | |
Line 379: | Line 417: | ||
|bgcolor="#F8F8F8"|'''Ligation:''' | |bgcolor="#F8F8F8"|'''Ligation:''' | ||
- | + | pLac-tetR | |
+ | |||
+ | |||
+ | pTet-mRFP | ||
+ | |||
|bgcolor="#E0E3FE"|'''Ligation:''' | |bgcolor="#E0E3FE"|'''Ligation:''' | ||
+ | |||
+ | pLac-TetR-ter | ||
|bgcolor="#CBD1FD"|'''Ligation:''' | |bgcolor="#CBD1FD"|'''Ligation:''' | ||
+ | |||
+ | pLac-ter-pTet-mRFP | ||
+ | |||
|bgcolor="#BCCDFD"|'''Ligation:''' | |bgcolor="#BCCDFD"|'''Ligation:''' | ||
+ | |||
+ | pLac-ter-pTet-mRFP-pSB3T5 | ||
+ | |||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
Line 392: | Line 442: | ||
|bgcolor="#E0E3FE"|'''Colony PCR :''' | |bgcolor="#E0E3FE"|'''Colony PCR :''' | ||
- | + | ok | |
|bgcolor="#CBD1FD"|'''Colony PCR:''' | |bgcolor="#CBD1FD"|'''Colony PCR:''' | ||
- | + | ok | |
|bgcolor="#BCCDFD"|'''Colony PCR :''' | |bgcolor="#BCCDFD"|'''Colony PCR :''' | ||
- | + | ok | |
|- 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:''' | ||
- | + | ok | |
|bgcolor="#CBD1FD"|'''Miniprep:''' | |bgcolor="#CBD1FD"|'''Miniprep:''' | ||
- | + | ok | |
|bgcolor="#BCCDFD"|'''Miniprep:''' | |bgcolor="#BCCDFD"|'''Miniprep:''' | ||
- | + | ok | |
|- 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 :''' | ||
- | + | ok | |
|bgcolor="#BCCDFD"|'''Sequencing :''' | |bgcolor="#BCCDFD"|'''Sequencing :''' | ||
- | + | ok | |
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
|bgcolor="#F8F8F8"| '''Stock glycerol:''' | |bgcolor="#F8F8F8"| '''Stock glycerol:''' | ||
Line 459: | Line 509: | ||
LB pure. | LB pure. | ||
+ | |||
+ | To learn more, see the [[Team:Paris/Conclusion | Conclusion page]] | ||
<html> | <html> | ||
Line 502: | Line 554: | ||
|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 513: | Line 570: | ||
ok | ok | ||
- | |bgcolor="#E0E3FE"| | + | |bgcolor="#E0E3FE"|- |
+ | |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"|''' | + | |bgcolor="#CBD1FD"|'''Digestion checking:''' |
- | + | none | |
- | |bgcolor="#BCCDFD"| | + | |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:''' | ||
+ | none | ||
- | |||
- | |||
|bgcolor="#CBD1FD"|'''Ligation:''' | |bgcolor="#CBD1FD"|'''Ligation:''' | ||
- | |bgcolor="#BCCDFD"| | + | none |
+ | |bgcolor="#BCCDFD"|- | ||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
Line 584: | Line 641: | ||
|bgcolor="#E0E3FE"|'''Colony PCR:''' | |bgcolor="#E0E3FE"|'''Colony PCR:''' | ||
- | + | none | |
|bgcolor="#CBD1FD"|'''Colony PCR:''' | |bgcolor="#CBD1FD"|'''Colony PCR:''' | ||
- | + | none | |
- | |bgcolor="#BCCDFD"| | + | |bgcolor="#BCCDFD"|- |
- | + | ||
- | - | + | |
|- 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 :''' | ||
- | + | none | |
|bgcolor="#CBD1FD"|'''Sequencing :''' | |bgcolor="#CBD1FD"|'''Sequencing :''' | ||
- | + | none | |
- | |bgcolor="#BCCDFD"| | + | |bgcolor="#BCCDFD"|- |
- | |||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
Line 632: | Line 684: | ||
|bgcolor="#E0E3FE"|'''Stock glycerol''' | |bgcolor="#E0E3FE"|'''Stock glycerol''' | ||
- | + | none | |
|bgcolor="#CBD1FD"|'''Stock glycerol''' | |bgcolor="#CBD1FD"|'''Stock glycerol''' | ||
- | + | none | |
- | |bgcolor="#BCCDFD"| | + | |bgcolor="#BCCDFD"|- |
- | + | ||
- | - | + | |
|} | |} | ||
Latest revision as of 03:58, 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
To learn more, see the Conclusion page
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.
To learn more, see the Conclusion page
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 | - |