Team:Paris/Addressing testing
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{{Template:Paris2009_menu3}} | {{Template:Paris2009_menu3}} | ||
- | |||
- | |||
- | |||
- | + | == WetLab - Addressing the message == | |
- | + | ||
<html> | <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_active" href="https://2009.igem.org/Team:Paris/Addressing_testing#bottom"> Addressing</a>| | ||
+ | <a class="menu_sub"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> | ||
- | <div id=" | + | <div id="right-side"></div> |
- | </div> | + | |
- | + | ||
</html> | </html> | ||
- | |||
- | ''' | + | |
+ | <font style="color:black;font-weight:bold; font-size:21px">'''Global constructions :'''</font> | ||
+ | |||
+ | |||
+ | |||
+ | <center><font style="color:green;font-weight:bold; font-size:16px">The green color means the experiment was a success</font></center> | ||
Line 26: | Line 81: | ||
- | |||
+ | |||
+ | <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; " | + | |- style="background: #0a3585; text-align: center; color:black; font-weight:bold;font-size:14px " |
- | |width="300px" ; bgcolor="#F8F8F8"| | + | |width="300px" ; bgcolor="#F8F8F8"| Column 1 |
- | |width="300px" ; bgcolor="#E0E3FE"| | + | |width="300px" ; bgcolor="#E0E3FE"| Column 2 |
- | |width="300px" ; bgcolor="#CBD1FD"| | + | |width="300px" ; bgcolor="#CBD1FD"| Column 3 |
- | |width="300px" ; bgcolor="#BCCDFD"| | + | |width="300px" ; bgcolor="#BCCDFD"| Column 4 |
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |'''PCR :''' | + | |bgcolor="#F8F8F8"|'''PCR :''' |
- | + | pBAD: plate 2008 | |
- | |||
- | + | ClyA Cterm matrix : [https://2009.igem.org/Team:Paris/Fridge F4] | |
- | + | Oligo :[https://2009.igem.org/Team:Paris/Freezer_Primers#O10 O10] and [https://2009.igem.org/Team:Paris/Freezer_Primers#O31 O31] TM :55°C | |
- | + | ||
- | |||
- | |||
- | + | mRFP Nterm plasmid : pSB1A3 | |
- | + | ||
+ | Oligo :O57 and O58 TM : | ||
+ | |||
+ | |||
+ | ClyA Nterm matrix : [https://2009.igem.org/Team:Paris/Fridge F4] | ||
+ | |||
+ | Oligo : [https://2009.igem.org/Team:Paris/Freezer_Primers#O32 O32] and [https://2009.igem.org/Team:Paris/Freezer_Primers#O9 O9] TM :55°C | ||
+ | |||
+ | |||
+ | mRFP Cterm matrix : pSB1A3 | ||
+ | |||
+ | Oligo :O59 and O60 TM : | ||
+ | |||
+ | |||
+ | |bgcolor="#E0E3FE"| | ||
+ | - | ||
+ | |bgcolor="#CBD1FD"| | ||
+ | - | ||
+ | |bgcolor="#BCCDFD"| | ||
- | - | ||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |'''Verification on gel :''' | + | |bgcolor="#F8F8F8"|'''Verification on gel :''' |
ok | ok | ||
- | | | + | |bgcolor="#E0E3FE"| |
- | + | ||
- | - | ||
- | | | + | |bgcolor="#CBD1FD"| |
- | - | ||
- | | | + | |bgcolor="#BCCDFD"| |
- | + | ||
- | - | ||
+ | |||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |'''Purification on gel :''' | + | |bgcolor="#F8F8F8"| '''Purification on gel :''' |
ok | ok | ||
- | | | + | |bgcolor="#E0E3FE"| |
- | + | ||
- | - | ||
- | |||
+ | |bgcolor="#CBD1FD"| | ||
- | - | ||
- | |||
+ | |bgcolor="#BCCDFD"| | ||
- | - | ||
+ | |||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |''' | + | |bgcolor="#F8F8F8"|'''Digestion:''' |
ClyA Cterm S/P | ClyA Cterm S/P | ||
+ | |||
RFP Nterm X/P | RFP Nterm X/P | ||
- | |||
+ | pBAD vector : ? E/S | ||
- | |||
- | + | |bgcolor="#E0E3FE"|'''Digestion:''' | |
- | + | ClyA Cter-RFP Nter vector PSB1A3 E/X | |
+ | |bgcolor="#CBD1FD"| | ||
- | - | ||
- | | | + | |bgcolor="#BCCDFD"| |
- | + | ||
- | - | ||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |''' | + | |bgcolor="#F8F8F8"| '''Verification digestion:''' |
ok | ok | ||
- | |''' | + | |bgcolor="#E0E3FE"|'''Verification digestion:''' |
ok | ok | ||
- | | | + | |bgcolor="#CBD1FD"|- |
- | + | |bgcolor="#BCCDFD"| | |
- | - | ||
- | |||
- | |||
- | |||
- | |||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |''' | + | |bgcolor="#F8F8F8"|'''Ligation:''' |
ClyA Cter-RFP Nter vector PSB1A3 | ClyA Cter-RFP Nter vector PSB1A3 | ||
- | |''' | + | |bgcolor="#E0E3FE"|'''Ligation:''' |
PBAD E/S ClyA Cter-RFP Nter PSB1A3 E/X (x2) | PBAD E/S ClyA Cter-RFP Nter PSB1A3 E/X (x2) | ||
- | |||
- | |''' | + | |
+ | PBAD S/P ClyA Cter-RFP Nter PSB2K3 X/P (x2) | ||
+ | |||
+ | |bgcolor="#CBD1FD"|'''Ligation:''' | ||
- | - | ||
- | | | + | |bgcolor="#BCCDFD"| |
- | + | ||
- | + | ||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |'''PCR | + | |bgcolor="#F8F8F8"| '''Colony PCR :''' |
ok | ok | ||
- | |'''PCR | + | |bgcolor="#E0E3FE"|'''Colony PCR :''' |
ok | ok | ||
- | |'''PCR | + | |bgcolor="#CBD1FD"|'''Colony PCR :''' |
- | + | ok | |
- | | | + | |bgcolor="#BCCDFD"| |
- | |||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |''' | + | |bgcolor="#F8F8F8"|'''Miniprep:''' |
clone : | clone : | ||
- | |''' | + | |bgcolor="#E0E3FE"|'''Miniprep:''' |
clone : | clone : | ||
- | |''' | + | |bgcolor="#CBD1FD"|'''Miniprep:''' |
- | + | clone 3 | |
- | | | + | |
+ | clone 5 | ||
+ | |||
+ | |bgcolor="#BCCDFD"| | ||
- | |||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |''' | + | |bgcolor="#F8F8F8"|'''Sequencing :''' |
ok | ok | ||
- | |''' | + | |bgcolor="#E0E3FE"|'''Sequencing :''' |
ok | ok | ||
- | |''' | + | |bgcolor="#CBD1FD"|'''Sequencing :''' |
- | + | ok | |
- | + | ||
+ | |bgcolor="#BCCDFD"| | ||
- | - | ||
+ | |||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |''' | + | |bgcolor="#F8F8F8"| '''Stock glycerol:''' |
- | + | ClyA CTer: S47(clone 3) and S48(clone 7) | |
- | + | ||
- | + | ClyA Nter: S72 | |
- | + | RFP Cter: S55 (Clone 3) | |
- | + | RFP Nter: S56 (Clone 3) | |
- | |||
- | |||
- | - | + | |bgcolor="#E0E3FE"|'''Stock glycerol''' |
+ | |||
+ | Cly A(Cter)-(Nter)RFP: S72(clone 8) | ||
+ | |||
+ | |bgcolor="#CBD1FD"|'''Stock glycerol''' | ||
+ | |||
+ | pBAD ClyA RFP : S89 (clone 3) | ||
+ | |||
+ | pBAD ClyA RFP: S90 (clone 5) | ||
+ | |||
+ | |bgcolor="#BCCDFD"| | ||
Line 227: | Line 300: | ||
- | PBAD ClyA RFP on PSB3T5 was transformed into Delta Tol bacteria , in this case we are supposed to see fluorescent vesicles | + | PBAD ClyA RFP on PSB3T5 was transformed into Delta Tol bacteria , in this case we are supposed to see fluorescent vesicles when the medium contains 1 % arabinose , and to have no fluorescent on 1% glucose. |
+ | |||
+ | |||
+ | To learn more, see the [[Team:Paris/Conclusion | Conclusion page]] | ||
Line 242: | Line 318: | ||
- | |||
+ | <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; " | + | |- style="background: #0a3585; text-align: center; color:black; font-weight:bold;font-size:14px " |
- | |width="300px" ; bgcolor="#F8F8F8"| | + | |width="300px" ; bgcolor="#F8F8F8"| Column 1 |
- | |width="300px" ; bgcolor="#E0E3FE"| | + | |width="300px" ; bgcolor="#E0E3FE"| Column 2 |
- | |width="300px" ; bgcolor="#CBD1FD"| | + | |width="300px" ; bgcolor="#CBD1FD"| Column 3 |
- | |width="300px" ; bgcolor="#BCCDFD"| | + | |width="300px" ; bgcolor="#BCCDFD"| Column 4 |
- | + | ||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |'''PCR :''' | + | |bgcolor="#F8F8F8"|'''PCR :''' |
TatABCE matrix : | TatABCE matrix : | ||
- | | | + | |bgcolor="#E0E3FE"|- |
- | | | + | |bgcolor="#CBD1FD"|- |
- | | | + | |bgcolor="#BCCDFD"|- |
+ | |||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |'''Verification on gel :''' | + | |bgcolor="#F8F8F8"|'''Verification on gel :''' |
ok | ok | ||
- | | | + | |bgcolor="#E0E3FE"|- |
- | + | ||
- | - | + | |
- | | | + | |bgcolor="#CBD1FD"| |
- | - | ||
- | | | + | |bgcolor="#BCCDFD"| |
- | - | ||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |'''Purification on gel :''' | + | |bgcolor="#F8F8F8"|'''Purification on gel :''' |
ok | ok | ||
- | | | + | |bgcolor="#E0E3FE"| |
- | - | ||
- | | | + | |bgcolor="#CBD1FD"| |
- | - | ||
- | | | + | |bgcolor="#BCCDFD"| |
- | - | ||
Line 301: | Line 374: | ||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |''' | + | |bgcolor="#F8F8F8"|'''Digestion:''' |
- | + | none | |
- | |''' | + | |bgcolor="#E0E3FE"|'''Digestion:''' |
- | |''' | + | none |
- | |''' | + | |bgcolor="#CBD1FD"|'''Digestion:''' |
+ | none | ||
+ | |bgcolor="#BCCDFD"|'''Digestion:''' | ||
+ | none | ||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |''' | + | |bgcolor="#F8F8F8"|'''Digestion checking:''' |
+ | none | ||
- | + | |bgcolor="#E0E3FE"|'''Digestion checking:''' | |
+ | none | ||
- | |||
- | + | |bgcolor="#CBD1FD"|'''Digestion checking:''' | |
- | |''' | + | none |
+ | |bgcolor="#BCCDFD"|'''Digestion checking:''' | ||
- | + | none | |
- | + | ||
- | + | ||
- | + | ||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |''' | + | |bgcolor="#F8F8F8"|'''Ligation:''' |
- | + | none | |
- | + | |bgcolor="#E0E3FE"|'''Ligation:''' | |
- | |''' | + | none |
- | |''' | + | |bgcolor="#CBD1FD"|'''Ligation:''' |
- | |''' | + | none |
+ | |bgcolor="#BCCDFD"|'''Ligation:''' | ||
+ | none | ||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |'''PCR | + | |bgcolor="#F8F8F8"|'''Colony PCR :''' |
+ | none | ||
- | + | |bgcolor="#E0E3FE"|'''Colony PCR :''' | |
+ | none | ||
- | |'''PCR | + | |bgcolor="#CBD1FD"|'''Colony PCR :''' |
+ | none | ||
+ | |bgcolor="#BCCDFD"|'''Colony PCR :''' | ||
+ | none | ||
- | - | + | |- style="background: #d8d8d8; text-align: center;" |
+ | |bgcolor="#F8F8F8"|'''Miniprep:''' | ||
- | + | none | |
- | + | |bgcolor="#E0E3FE"|'''Miniprep:''' | |
- | |''' | + | |
- | + | none | |
- | | | + | |bgcolor="#CBD1FD"|'''Miniprep:''' |
- | |''' | + | |
- | + | none | |
- | |''' | + | |bgcolor="#BCCDFD"|'''Miniprep:''' |
- | + | none | |
- | |''' | + | |- style="background: #bebebe; text-align: center;" |
+ | |bgcolor="#F8F8F8"|'''Sequencing :''' | ||
- | + | none | |
- | |''' | + | |bgcolor="#E0E3FE"|'''Sequencing :''' |
- | + | none | |
- | | | + | |bgcolor="#CBD1FD"|'''Sequencing :''' |
- | |''' | + | |
- | + | none | |
+ | |bgcolor="#BCCDFD"|'''Sequencing :''' | ||
- | + | none | |
- | - | + | |- style="background: #d8d8d8; text-align: center;" |
+ | |bgcolor="#F8F8F8"| '''Stock glycerol:''' | ||
- | |''' | + | none |
+ | |bgcolor="#E0E3FE"|'''Stock glycerol''' | ||
- | + | none | |
- | |''' | + | |bgcolor="#CBD1FD"|'''Stock glycerol''' |
- | + | none | |
+ | |bgcolor="#BCCDFD"|'''Stock glycerol''' | ||
+ | |||
+ | none | ||
|} | |} | ||
- | + | <html> | |
+ | <a href="https://2009.igem.org/Team:Paris/Addressing_testing#bottom"><img style="width:40px; height:40px;" src="https://static.igem.org/mediawiki/2009/1/10/Paris_Up.png"/></a> | ||
+ | </html> |
Latest revision as of 03:53, 22 October 2009
iGEM > Paris > WetLab > Addressing
WetLab - Addressing the message
Global constructions :
Experiments ran :
Column 1 | Column 2 | Column 3 | Column 4 |
PCR :
pBAD: plate 2008
Oligo :O57 and O58 TM :
Oligo :O59 and O60 TM :
|
- |
- |
- |
Verification on gel :
ok |
- |
- |
-
|
Purification on gel :
ok |
-
|
- |
-
|
Digestion:
ClyA Cterm S/P
| Digestion:
ClyA Cter-RFP Nter vector PSB1A3 E/X |
- |
- |
Verification digestion:
ok | Verification digestion:
ok | - |
- |
Ligation:
ClyA Cter-RFP Nter vector PSB1A3 | Ligation:
PBAD E/S ClyA Cter-RFP Nter PSB1A3 E/X (x2)
| Ligation:
- |
|
Colony PCR :
ok | Colony PCR :
ok | Colony PCR :
ok |
|
Miniprep:
clone : | Miniprep:
clone : | Miniprep:
clone 3 clone 5 |
|
Sequencing :
ok | Sequencing :
ok | Sequencing :
ok |
-
|
Stock glycerol:
ClyA CTer: S47(clone 3) and S48(clone 7) ClyA Nter: S72 RFP Cter: S55 (Clone 3) RFP Nter: S56 (Clone 3)
| Stock glycerol
Cly A(Cter)-(Nter)RFP: S72(clone 8) | Stock glycerol
pBAD ClyA RFP : S89 (clone 3) pBAD ClyA RFP: S90 (clone 5) |
|
Functional Testing:
PBAD ClyA RFP was transformed into Top10 bacteria in order to localize the fluorescence, we are supposed to have a superior fluorescence in the membrane.
PBAD ClyA RFP on PSB3T5 was transformed into Delta Tol bacteria , in this case we are supposed to see fluorescent vesicles when the medium contains 1 % arabinose , and to have no fluorescent on 1% glucose.
To learn more, see the Conclusion page
Export system
We finally thought that it won't be neccesary to overexpress the Tat system, nevertheless we have run a few experiments before starting to focus on others parts of the project.
Experiments ran :
Column 1 | Column 2 | Column 3 | Column 4 |
PCR :
TatABCE matrix : | - | - | -
|
Verification on gel :
ok | - |
- |
- |
Purification on gel :
ok |
- |
- |
-
|
Digestion:
none | Digestion:
none | Digestion:
none | Digestion:
none |
Digestion checking:
none | Digestion checking:
none
| Digestion checking:
none | Digestion checking:
none
|
Ligation:
none | Ligation:
none | Ligation:
none | Ligation:
none |
Colony PCR :
none | Colony PCR :
none | Colony PCR :
none | Colony PCR :
none |
Miniprep:
none | Miniprep:
none | Miniprep:
none | Miniprep:
none |
Sequencing :
none | Sequencing :
none | Sequencing :
none | Sequencing :
none |
Stock glycerol:
none | Stock glycerol
none | Stock glycerol
none | Stock glycerol
none |