Team:Paris/Production testing

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Latest revision as of 03:58, 22 October 2009

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WetLab - Production

Main | Addressing| Production| Reception

The delay

We separate the construction on two plasmids : Part I and Part II.

Part I : Testing delay phase I

The green color means the experiment was a success

Experiments ran :

Column 1	Column 2	Column 3	Column 4
PCR : pBAD: plate igem LacI: plate igem Terminator: plate igem RBS: plate igem YFP: 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	-
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

The green color means the experiment was a success

Experiments ran :

Column 1	Column 2	Column 3	Column 4
PCR : terminator: igem plate pLac: igem plate TetR pTet: mRFP	-	-	-
Verification on gel : ok	- -	- -	- -
Purification on gel : ok	-	-	-
Digestion: pLac S/P TetR X/P terminator E/X pTet S/P mRFP X/P pSB3T5 E/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 pTet-mRFP	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

The green color means the experiment was a success

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 TolRII X/P g3p 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	-

@@ Line 1: / Line 1: @@
-<span/ id="bottom">[https://2009.igem.org/ iGEM ] > [[Team:Paris#top | Paris]] > [[Team:Paris/Production_overview#top | Production]] > [[Team:Paris/Production_testing#bottom | Testing]]
+<span/ id="bottom">[https://2009.igem.org/ iGEM ] > [[Team:Paris#top | Paris]] > [[team:Paris/WetLab#WetLab| WetLab]] > [[Team:Paris/Production_testing#bottom | Production]]
 {{Template:Paris2009}}
 {{Template:Paris2009_menu2}}
-== WetLab ==
-<center> [[team:Paris/WetLab#WetLab| Main]] - [[Team:Paris/Addressing_testing#top|Addressing]] - [[Team:Paris/Production_testing#top| Production]] - [[Team:Paris/Transduction_testing#top | reception]] - </center>
+== WetLab - Production ==
-<center> '''Production'''</center>
+<html>
+<style type="text/css">
+#left-side {
+    position: absolute;
+    height: 23px;
+    width: 30px;
+    top: 0px;
+    left: 180px;
+    margin-top:10px;
+    padding-top: 7px;
+    background: url(https://static.igem.org/mediawiki/2009/1/1b/Left_menu_pari.png);
+    z-index:4;
+}
+#middle-side {
+    height: 25px;
+    width: 320px;
+    position: absolute;
+    top: 0px;
+    left: 190px;
+    margin-top:10px;
+    padding-top: 5px;
+    background: #dadada;
+    z-index:5;
+}
+#right-side {
+    position: absolute;
+    height: 23px;
+    width: 30px;
+    margin-top:10px;
+    padding-top: 7px;
+    top: 0px;
+    left: 490px;
+    background: url(https://static.igem.org/mediawiki/2009/4/40/Right_menu_paris.png);
+    z-index:4;
+}
+a.menu_sub {
+    padding-left: 7px;
+    padding-right: 7px;
+}
+a.menu_sub_active {
+    padding-left: 7px;
+    padding-right: 7px;
+    color:#b0310e;
+    font-weight:bold;
+}
+</style>
+<div id="left-side"></div>
+<div id="middle-side"><center>
+<a class="menu_sub"href="https://2009.igem.org/Team:Paris/WetLab#bottom"> Main </a>|
+<a class="menu_sub" href="https://2009.igem.org/Team:Paris/Addressing_testing#bottom"> Addressing</a>|
+<a class="menu_sub_active"href="https://2009.igem.org/Team:Paris/Production_testing#bottom"> Production</a>|
+<a class="menu_sub"href="https://2009.igem.org/Team:Paris/Transduction_testing#bottom"> Reception</a>
+</center>
+</div>
+<div id="right-side"></div>
+</html>
+#[[Team:Paris/Production_testing#The delay | The delay]]
+##[[Team:Paris/Production_testing#Part I : Testing delay phase I| Part I : Testing delay phase I]]
+##[[Team:Paris/Production_testing#Part I : Functional testing of the part I| Part I : Functional testing of the part I]]
+##[[Team:Paris/Production_testing#Part II : Testing delay phase II| Part II : Testing delay phase II]]
+##[[Team:Paris/Production_testing#Part II : Functional testing of the part II| Part II : Functional testing of the part II]]
+#[[Team:Paris/Production_testing#The destabilization of the membrane| The destabilization of the membrane]]
 <html>
 </div>
@@ Line 13: / Line 80: @@
 </html>
-== tests ==
+==The delay ==
-two main construction :
+We separate the construction on two plasmids : Part I and Part II.
-the delay
+===Part I : Testing delay phase I===
-the destabilization of the membrane
+<center><font style="color:green;font-weight:bold; font-size:16px">The green color means the experiment was a success</font></center>
+[[Image:Paris_construc_3_delay1.jpg  |800px|center|plasmid = PSB3T5]]
+<font style="color:black;font-weight:bold; font-size:18px"><I>'''Experiments ran :'''</I></font>
+{|
+|- style="background: #0a3585; text-align: center; color:black; font-weight:bold;font-size:14px "
+|width="300px" ; bgcolor="#F8F8F8"| Column 1
+|width="300px" ; bgcolor="#E0E3FE"| Column 2
+|width="300px" ; bgcolor="#CBD1FD"| Column 3
+|width="300px" ; bgcolor="#BCCDFD"| Column 4
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''PCR :'''
+pBAD: plate igem
+LacI: plate igem
+Terminator: plate igem
+RBS: plate igem
+YFP: plate igem
+|bgcolor="#E0E3FE"|-
+|bgcolor="#CBD1FD"|-
+|bgcolor="#BCCDFD"|-
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"|'''Verification on gel :'''
+ok
+|bgcolor="#E0E3FE"|-
+|bgcolor="#CBD1FD"|-
+|bgcolor="#BCCDFD"|-
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''Purification on gel :'''
+ok
+|bgcolor="#E0E3FE"|-
+|bgcolor="#CBD1FD"|-
+|bgcolor="#BCCDFD"|-
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"|'''Digestion:'''
+pBAD S/P
+lacI E/S
+terminator E/X
+RBS S/P
+YFP X/P
+|bgcolor="#E0E3FE"|'''Digestion:'''
+pBAD-LacI  E/S
+RBS-YFP  E/S
+|bgcolor="#CBD1FD"|'''Digestion:'''
+RBS-YFP-ter  E/X
+|bgcolor="#BCCDFD"|-
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''Verification digestion:'''
+ok
+|bgcolor="#E0E3FE"|'''Verification digestion:'''
+ok
+|bgcolor="#CBD1FD"|'''Verification digestion:'''
+ok
+|bgcolor="#BCCDFD"|-
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"|'''Ligation:'''
+pBAD-LacI
+RBS-YFP
+|bgcolor="#E0E3FE"|'''Ligation:'''
+|bgcolor="#CBD1FD"|'''Ligation:'''
+|bgcolor="#BCCDFD"|'''Ligation:'''
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''Colony PCR :'''
+ok
+|bgcolor="#E0E3FE"|'''Colony PCR :'''
+ok
+|bgcolor="#CBD1FD"|'''Colony PCR :'''
+ok
+|bgcolor="#BCCDFD"|'''Colony PCR:'''
+ok
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"|'''Miniprep:'''
+clone
+|bgcolor="#E0E3FE"|'''Miniprep:'''
+clone
+|bgcolor="#CBD1FD"|'''Miniprep:'''
+clone
+|bgcolor="#BCCDFD"|'''Miniprep:'''
+clone
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''Sequencing :'''
+ok
+|bgcolor="#E0E3FE"|'''Sequencing :'''
+ok
+|bgcolor="#CBD1FD"|'''Sequencing :'''
+ok
+|bgcolor="#BCCDFD"|'''Sequencing :'''
+ok
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"| '''Stock glycerol:'''
+|bgcolor="#E0E3FE"|'''Stock glycerol'''
+Cly A(Cter)-(Nter)RFP: S72(clone 8)
+|bgcolor="#CBD1FD"|'''Stock glycerol'''
+-
+|bgcolor="#BCCDFD"|'''Stock glycerol'''
+-
+|}
+===Part I : Functional testing of the part I===
+PBAD LacI LVA- YFP was transforme into Top ten Bacteria and grew on medium :
+% arabinose
+% glucose
+LB pure
+To learn more, see the [[Team:Paris/Conclusion | Conclusion page]]
+<html>
+<a href="https://2009.igem.org/Team:Paris/Production_testing#bottom"><img style="width:40px; height:40px;" src="https://static.igem.org/mediawiki/2009/1/10/Paris_Up.png"/></a>
+</html>
 <html>
@@ Line 28: / Line 285: @@
 </html>
+===Part II : Testing delay phase II===
-== delay ==
-part1 :
-[[Image:Paris_construc_3_delay1.jpg  |700px|thumb|left|plasmid = PSB3T5]]
+<center><font style="color:green;font-weight:bold; font-size:16px">The green color means the experiment was a success</font></center>
-part2 :
+[[Image:Paris_construc_5_delay2.jpg  |800px|center|plasmid = ]]
-[[Image:Paris_construc_5_delay2.jpg  |700px|thumb|left|plasmid = PSB3T5]]
+<font style="color:black;font-weight:bold; font-size:18px"><I>'''Experiments ran :'''</I></font>
+{|
+|- style="background: #0a3585; text-align: center; color:black; font-weight:bold;font-size:14px "
+|width="300px" ; bgcolor="#F8F8F8"| Column 1
+|width="300px" ; bgcolor="#E0E3FE"| Column 2
+|width="300px" ; bgcolor="#CBD1FD"| Column 3
+|width="300px" ; bgcolor="#BCCDFD"| Column 4
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''PCR :'''
+terminator: igem plate
+pLac: igem plate
+TetR
+pTet:
+mRFP
+|bgcolor="#E0E3FE"|-
+|bgcolor="#CBD1FD"|-
+|bgcolor="#BCCDFD"|-
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"|'''Verification on gel :'''
+ok
+|bgcolor="#E0E3FE"|-
+-
+|bgcolor="#CBD1FD"|-
+-
+|bgcolor="#BCCDFD"|-
+-
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''Purification on gel :'''
+ok
+|bgcolor="#E0E3FE"|
+-
+|bgcolor="#CBD1FD"|
+-
+|bgcolor="#BCCDFD"|
+-
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"|'''Digestion:'''
+pLac  S/P
+TetR X/P
+terminator  E/X
+pTet  S/P
+mRFP  X/P
+pSB3T5  E/P
+|bgcolor="#E0E3FE"|'''Digestion:'''
+plac-TetR  E/S
+pTet-mRFP  X/P
+|bgcolor="#CBD1FD"|'''Digestion:'''
+pLac-tetR-ter  S/P
+|bgcolor="#BCCDFD"|'''Digestion:'''
+pLac-tetR-ter-pTet-mRFP  E/P
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''Digestion checking:'''
+ok
+|bgcolor="#E0E3FE"|'''Digestion checking:'''
+ok
+|bgcolor="#CBD1FD"|'''Digestion checking:'''
+ok
+|bgcolor="#BCCDFD"|'''Digestion checking:'''
+ok
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"|'''Ligation:'''
+pLac-tetR
+pTet-mRFP
+|bgcolor="#E0E3FE"|'''Ligation:'''
+pLac-TetR-ter
+|bgcolor="#CBD1FD"|'''Ligation:'''
+pLac-ter-pTet-mRFP
+|bgcolor="#BCCDFD"|'''Ligation:'''
+pLac-ter-pTet-mRFP-pSB3T5
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''Colony PCR:'''
+ok
+|bgcolor="#E0E3FE"|'''Colony PCR :'''
+ok
+|bgcolor="#CBD1FD"|'''Colony PCR:'''
+ok
+|bgcolor="#BCCDFD"|'''Colony PCR :'''
+ok
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"|'''Miniprep:'''
+ok
+|bgcolor="#E0E3FE"|'''Miniprep:'''
+ok
+|bgcolor="#CBD1FD"|'''Miniprep:'''
+ok
+|bgcolor="#BCCDFD"|'''Miniprep:'''
+ok
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''Sequencing :'''
+ok
+|bgcolor="#E0E3FE"|'''Sequencing :'''
+ok
+|bgcolor="#CBD1FD"|'''Sequencing :'''
+ok
+|bgcolor="#BCCDFD"|'''Sequencing :'''
+ok
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"| '''Stock glycerol:'''
+-
+|bgcolor="#E0E3FE"|'''Stock glycerol'''
+-
+|bgcolor="#CBD1FD"|'''Stock glycerol'''
+-
+|bgcolor="#BCCDFD"|'''Stock glycerol'''
+-
+|}
+===Part II : Functional testing of the part II===
+Ptet RFP Plac TetR was tranforme into Top10 bacteria and grew on media :
+,1% IPTG .
+% glucose.
+LB pure.
+To learn more, see the [[Team:Paris/Conclusion | Conclusion page]]
+<html>
+<a href="https://2009.igem.org/Team:Paris/Production_testing#bottom"><img style="width:40px; height:40px;" src="https://static.igem.org/mediawiki/2009/1/10/Paris_Up.png"/></a>
+</html>
@@ Line 48: / Line 524: @@
 </html>
-== destabilization ==
+==The destabilization of the membrane==
-<b class="correction">
-On rentre dans les détails de nos tests & résultats :
-* On a X temps disponible devant nous pour une équipe de X personnes 'dédiées' a cette partie (liste des coco qui s'en occupent ... permet de personnalisé un peu et de rendre plus vivant).
-* On découpe dans les étapes AVEC les étapes de vérifications et constructions de celles-ci depuis le début le tout en un listing un peu détaillé avec le plus de liens possible (freezer, protocols, notebook etc...). Exemple :
-** on sait qu'il existe des BB, on les vérifies
-** on demande a nos pote de nous filer leur constructions (ou pas)
+<center><font style="color:green;font-weight:bold; font-size:16px">The green color means the experiment was a success</font></center>
-** on entame la vérification de ca : renvois vers la construction bidule
-** ok, on passe à la suivante etc etc..
-</b>
+[[Image:Paris_construc_4_tr2.jpg |800px|center|plasmid = PSB3T5]]
+<font style="color:black;font-weight:bold; font-size:18px"><I>'''Experiments ran :'''</I></font>
+{|
+|- style="background: #0a3585; text-align: center; color:black; font-weight:bold;font-size:14px "
+|width="300px" ; bgcolor="#F8F8F8"| Column 1
+|width="300px" ; bgcolor="#E0E3FE"| Column 2
+|width="300px" ; bgcolor="#CBD1FD"| Column 3
+|width="300px" ; bgcolor="#BCCDFD"| Column 4
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''PCR :'''
+pTet: igem plate
+OmpAs:
+ToRII: pINK2
+g3p: pINTg3p
+|bgcolor="#E0E3FE"|
+|bgcolor="#CBD1FD"|'
+|bgcolor="#BCCDFD"|
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"|'''Verification on gel :'''
+ok
+|bgcolor="#E0E3FE"|-
+|bgcolor="#CBD1FD"|-
+|bgcolor="#BCCDFD"|-
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''Purification on gel :'''
+ok
+|bgcolor="#E0E3FE"|-
+|bgcolor="#CBD1FD"|-
+|bgcolor="#BCCDFD"|-
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"|'''Digestion:'''
+pTet  S/P
+OmpAs  X/P
+TolRII   X/P
+g3p   X/P
+|bgcolor="#E0E3FE"|'''Digestion:'''
+pTet-OmpAs  S/P
+|bgcolor="#CBD1FD"|'''Digestion:'''
+none
+|bgcolor="#BCCDFD"|-
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''Digestion checking:'''
+ok
+|bgcolor="#E0E3FE"|'''Digestion checking:'''
+ok
+|bgcolor="#CBD1FD"|'''Digestion checking:'''
+none
+|bgcolor="#BCCDFD"|-
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"|'''Ligation:'''
+pTet-OmpAs
+|bgcolor="#E0E3FE"|'''Ligation:'''
+none
+|bgcolor="#CBD1FD"|'''Ligation:'''
+none
+|bgcolor="#BCCDFD"|-
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''Colony PCR :'''
+ok
+|bgcolor="#E0E3FE"|'''Colony PCR:'''
+none
+|bgcolor="#CBD1FD"|'''Colony PCR:'''
+none
+|bgcolor="#BCCDFD"|-
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"|'''Miniprep:'''
+ok
+|bgcolor="#E0E3FE"|'''Miniprep:'''
+none
+|bgcolor="#CBD1FD"|'''Miniprep:'''
+none
+|bgcolor="#BCCDFD"|-
+|- style="background: #bebebe; text-align: center;"
+|bgcolor="#F8F8F8"|'''Sequencing :'''
+ok
+|bgcolor="#E0E3FE"|'''Sequencing :'''
+none
+|bgcolor="#CBD1FD"|'''Sequencing :'''
+none
+|bgcolor="#BCCDFD"|-
+|- style="background: #d8d8d8; text-align: center;"
+|bgcolor="#F8F8F8"| '''Stock glycerol:'''
+-
+|bgcolor="#E0E3FE"|'''Stock glycerol'''
+none
+|bgcolor="#CBD1FD"|'''Stock glycerol'''
+none
+|bgcolor="#BCCDFD"|-
+|}
+<html>
+<a href="https://2009.igem.org/Team:Paris/Production_testing#bottom"><img style="width:40px; height:40px;" src="https://static.igem.org/mediawiki/2009/1/10/Paris_Up.png"/></a>
+</html>

Team:Paris/Production testing

From 2009.igem.org

Latest revision as of 03:58, 22 October 2009

Contents

WetLab - Production

The delay

Part I : Testing delay phase I

Part I : Functional testing of the part I

Part II : Testing delay phase II

Part II : Functional testing of the part II

The destabilization of the membrane