Team:Paris/Addressing testing
From 2009.igem.org
(→WetLab - Addressing the message) |
(→Functional Testing:) |
||
(43 intermediate revisions not shown) | |||
Line 69: | Line 69: | ||
- | |||
- | <center><font style="color: | + | <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> | ||
+ | |||
[[Image:Paris_construc_1_PCR.jpg |800px|center|plasmid = PSB3T5]] | [[Image:Paris_construc_1_PCR.jpg |800px|center|plasmid = PSB3T5]] | ||
Line 77: | 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;font-size:14px " | |- 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;" | ||
Line 117: | Line 121: | ||
- | |bgcolor="#E0E3FE"| | + | |bgcolor="#E0E3FE"| |
- | + | ||
- | - | ||
- | |bgcolor="#CBD1FD"| | + | |bgcolor="#CBD1FD"| |
- | + | ||
- | - | ||
- | |bgcolor="#BCCDFD"| | + | |bgcolor="#BCCDFD"| |
- | + | ||
- | - | ||
Line 132: | Line 133: | ||
ok | ok | ||
- | |bgcolor="#E0E3FE"| | + | |bgcolor="#E0E3FE"| |
- | + | ||
- | - | ||
- | |bgcolor="#CBD1FD"| | + | |bgcolor="#CBD1FD"| |
- | - | ||
- | |bgcolor="#BCCDFD"| | + | |bgcolor="#BCCDFD"| |
- | + | ||
- | - | ||
+ | |||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
Line 149: | Line 149: | ||
ok | ok | ||
- | |bgcolor="#E0E3FE"| | + | |bgcolor="#E0E3FE"| |
- | + | ||
- | - | ||
- | |||
+ | |bgcolor="#CBD1FD"| | ||
- | - | ||
- | |||
+ | |bgcolor="#BCCDFD"| | ||
- | - | ||
+ | |||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
Line 177: | Line 177: | ||
ClyA Cter-RFP Nter vector PSB1A3 E/X | ClyA Cter-RFP Nter vector PSB1A3 E/X | ||
- | |bgcolor="#CBD1FD"| | + | |bgcolor="#CBD1FD"| |
- | + | ||
- | - | ||
- | |bgcolor="#BCCDFD"| | + | |bgcolor="#BCCDFD"| |
- | + | ||
- | - | ||
Line 193: | Line 191: | ||
ok | ok | ||
- | |bgcolor="#CBD1FD"| | + | |bgcolor="#CBD1FD"|- |
- | + | |bgcolor="#BCCDFD"| | |
- | - | ||
- | |||
- | |||
- | |||
- | |||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
Line 216: | Line 210: | ||
- | - | ||
- | |bgcolor="#BCCDFD"| | + | |bgcolor="#BCCDFD"| |
- | + | ||
- | + | ||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |bgcolor="#F8F8F8"| '''PCR | + | |bgcolor="#F8F8F8"| '''Colony PCR :''' |
ok | ok | ||
Line 231: | Line 224: | ||
|bgcolor="#CBD1FD"|'''Colony PCR :''' | |bgcolor="#CBD1FD"|'''Colony PCR :''' | ||
- | + | ok | |
- | |bgcolor="#BCCDFD"| | + | |bgcolor="#BCCDFD"| |
- | |||
Line 249: | Line 241: | ||
|bgcolor="#CBD1FD"|'''Miniprep:''' | |bgcolor="#CBD1FD"|'''Miniprep:''' | ||
- | + | clone 3 | |
- | |bgcolor="#BCCDFD"| | + | |
+ | clone 5 | ||
+ | |||
+ | |bgcolor="#BCCDFD"| | ||
- | |||
Line 266: | Line 260: | ||
|bgcolor="#CBD1FD"|'''Sequencing :''' | |bgcolor="#CBD1FD"|'''Sequencing :''' | ||
- | + | ok | |
- | + | ||
+ | |bgcolor="#BCCDFD"| | ||
- | - | ||
+ | |||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
Line 289: | Line 284: | ||
|bgcolor="#CBD1FD"|'''Stock glycerol''' | |bgcolor="#CBD1FD"|'''Stock glycerol''' | ||
- | + | pBAD ClyA RFP : S89 (clone 3) | |
- | + | ||
- | + | pBAD ClyA RFP: S90 (clone 5) | |
+ | |||
+ | |bgcolor="#BCCDFD"| | ||
Line 304: | 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 319: | 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;" | ||
Line 339: | Line 336: | ||
TatABCE matrix : | TatABCE matrix : | ||
- | |bgcolor="#E0E3FE"| | + | |bgcolor="#E0E3FE"|- |
- | |bgcolor="#CBD1FD"| | + | |bgcolor="#CBD1FD"|- |
- | |bgcolor="#BCCDFD"| | + | |bgcolor="#BCCDFD"|- |
+ | |||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
Line 348: | Line 346: | ||
ok | ok | ||
- | |bgcolor="#E0E3FE"| | + | |bgcolor="#E0E3FE"|- |
- | + | ||
- | - | + | |
- | |bgcolor="#CBD1FD"| | + | |bgcolor="#CBD1FD"| |
- | - | ||
- | |bgcolor="#BCCDFD"| | + | |bgcolor="#BCCDFD"| |
- | - | ||
Line 364: | Line 360: | ||
ok | ok | ||
- | |bgcolor="#E0E3FE"| | + | |bgcolor="#E0E3FE"| |
- | - | ||
- | |bgcolor="#CBD1FD"| | + | |bgcolor="#CBD1FD"| |
- | - | ||
- | |bgcolor="#BCCDFD"| | + | |bgcolor="#BCCDFD"| |
- | - | ||
Line 378: | Line 374: | ||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |bgcolor="#F8F8F8"|''' | + | |bgcolor="#F8F8F8"|'''Digestion:''' |
- | + | none | |
- | |bgcolor="#E0E3FE"|''' | + | |bgcolor="#E0E3FE"|'''Digestion:''' |
- | |bgcolor="#CBD1FD"|''' | + | none |
- | |bgcolor="#BCCDFD"|''' | + | |bgcolor="#CBD1FD"|'''Digestion:''' |
+ | none | ||
+ | |bgcolor="#BCCDFD"|'''Digestion:''' | ||
+ | none | ||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |bgcolor="#F8F8F8"|''' | + | |bgcolor="#F8F8F8"|'''Digestion checking:''' |
+ | none | ||
- | + | |bgcolor="#E0E3FE"|'''Digestion checking:''' | |
+ | none | ||
- | |||
- | + | |bgcolor="#CBD1FD"|'''Digestion checking:''' | |
- | |bgcolor="# | + | none |
+ | |bgcolor="#BCCDFD"|'''Digestion checking:''' | ||
- | + | none | |
- | + | ||
- | + | ||
- | + | ||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |bgcolor="#F8F8F8"|''' | + | |bgcolor="#F8F8F8"|'''Ligation:''' |
- | + | none | |
- | + | |bgcolor="#E0E3FE"|'''Ligation:''' | |
- | |bgcolor="#E0E3FE"|''' | + | none |
- | |bgcolor="#CBD1FD"|''' | + | |bgcolor="#CBD1FD"|'''Ligation:''' |
- | |bgcolor="#BCCDFD"|''' | + | none |
+ | |bgcolor="#BCCDFD"|'''Ligation:''' | ||
+ | none | ||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |bgcolor="#F8F8F8"|'''PCR | + | |bgcolor="#F8F8F8"|'''Colony PCR :''' |
+ | none | ||
- | + | |bgcolor="#E0E3FE"|'''Colony PCR :''' | |
+ | none | ||
- | + | |bgcolor="#CBD1FD"|'''Colony PCR :''' | |
- | + | none | |
- | + | |bgcolor="#BCCDFD"|'''Colony PCR :''' | |
- | + | none | |
- | |bgcolor="#CBD1FD"|'''PCR | + | |
- | + | ||
- | + | ||
- | |bgcolor="#BCCDFD"|'''PCR | + | |
- | + | ||
- | + | ||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |bgcolor="#F8F8F8"|''' | + | |bgcolor="#F8F8F8"|'''Miniprep:''' |
- | + | none | |
- | |bgcolor="#E0E3FE"|''' | + | |bgcolor="#E0E3FE"|'''Miniprep:''' |
- | + | none | |
- | |bgcolor="#CBD1FD"|''' | + | |bgcolor="#CBD1FD"|'''Miniprep:''' |
- | + | none | |
- | |bgcolor="#BCCDFD"|''' | + | |bgcolor="#BCCDFD"|'''Miniprep:''' |
- | + | none | |
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |bgcolor="#F8F8F8"|''' | + | |bgcolor="#F8F8F8"|'''Sequencing :''' |
- | + | none | |
- | |bgcolor="#E0E3FE"|''' | + | |bgcolor="#E0E3FE"|'''Sequencing :''' |
- | + | none | |
- | |bgcolor="#CBD1FD"|''' | + | |bgcolor="#CBD1FD"|'''Sequencing :''' |
- | + | none | |
- | |bgcolor="#BCCDFD"|''' | + | |bgcolor="#BCCDFD"|'''Sequencing :''' |
- | + | none | |
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |bgcolor="#F8F8F8"| ''' | + | |bgcolor="#F8F8F8"| '''Stock glycerol:''' |
- | + | none | |
- | |bgcolor="#E0E3FE"|''' | + | |bgcolor="#E0E3FE"|'''Stock glycerol''' |
- | + | none | |
- | |bgcolor="#CBD1FD"|''' | + | |bgcolor="#CBD1FD"|'''Stock glycerol''' |
- | + | none | |
- | |bgcolor="#BCCDFD"|''' | + | |bgcolor="#BCCDFD"|'''Stock glycerol''' |
- | + | none | |
|} | |} |
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 |