Team:Paris/Production testing
From 2009.igem.org
(→Part II : Testing delay phase II) |
(→The destabilization of the membrane) |
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{| | {| | ||
- | |- style="background: #0a3585; text-align: center; color: | + | |- style="background: #0a3585; text-align: center; color:black; font-weight:bold; " |
- | |width= 300px| | + | |width="300px" ; bgcolor="#F8F8F8"| column 1 |
- | |width=300px| | + | |width="300px" ; bgcolor="#E0E3FE"| column 2 |
- | |width=300px| | + | |width="300px" ; bgcolor="#CBD1FD"| column 3 |
+ | |width="300px" ; bgcolor="#BCCDFD"| column 4 | ||
+ | |||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |'''PCR :''' | + | |bgcolor="#F8F8F8"|'''PCR :''' |
- | + | ||
- | + | ||
- | + | ||
- | + | TatABCE matrix : | |
- | + | ||
+ | |bgcolor="#E0E3FE"|'''PCR :''' | ||
+ | |bgcolor="#CBD1FD"|'''PCR :''' | ||
+ | |bgcolor="#BCCDFD"|'''PCR :''' | ||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |'''Verification on gel :''' | + | |bgcolor="#F8F8F8"|'''Verification on gel :''' |
ok | ok | ||
- | |'''Verification on gel :''' | + | |bgcolor="#E0E3FE"|'''Verification on gel :''' |
- | + | - | |
- | |'''Verification on gel :''' | + | |bgcolor="#CBD1FD"|'''Verification on gel :''' |
- | + | - | |
+ | |bgcolor="#BCCDFD"|'''Verification on gel :''' | ||
+ | |||
+ | - | ||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |'''Purification on gel :''' | + | |bgcolor="#F8F8F8"|'''Purification on gel :''' |
ok | ok | ||
- | |'''Purification on gel :''' | + | |bgcolor="#E0E3FE"|'''Purification on gel :''' |
- | + | - | |
- | |'''Purification on gel :''' | + | |bgcolor="#CBD1FD"|'''Purification on gel :''' |
- | + | - | |
+ | |bgcolor="#BCCDFD"|'''Purification on gel :''' | ||
+ | - | ||
- | |||
- | |||
+ | |- style="background: #d8d8d8; text-align: center;" | ||
+ | |bgcolor="#F8F8F8"|'''digestion:''' | ||
+ | TatABCE | ||
- | + | |bgcolor="#E0E3FE"|'''digestion:''' | |
- | |'''digestion:''' | + | |bgcolor="#CBD1FD"|'''digestion:''' |
- | |'''digestion:''' | + | |bgcolor="#BCCDFD"|'''digestion:''' |
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |'''verification digestion:''' | + | |bgcolor="#F8F8F8"|'''verification digestion:''' |
ok | ok | ||
- | |'''verification digestion:''' | + | |bgcolor="#E0E3FE"|'''verification digestion:''' |
- | + | - | |
- | |'''verification digestion:''' | + | |bgcolor="#CBD1FD"|'''verification digestion:''' |
- | + | - | |
+ | |bgcolor="#BCCDFD"|'''verification digestion:''' | ||
+ | |||
+ | - | ||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |'''ligation:''' | + | |bgcolor="#F8F8F8"|'''ligation:''' |
- | + | ||
- | + | ||
- | + | TatABCE | |
- | + | ||
+ | |bgcolor="#E0E3FE"|'''ligation:''' | ||
+ | |bgcolor="#CBD1FD"|'''ligation:''' | ||
+ | |bgcolor="#BCCDFD"|'''ligation:''' | ||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |'''PCR colo :''' | + | |bgcolor="#F8F8F8"|'''PCR colo :''' |
ok | ok | ||
- | |'''PCR colo :''' | + | |bgcolor="#E0E3FE"|'''PCR colo :''' |
- | + | - | |
- | |'''PCR colo :''' | + | |bgcolor="#CBD1FD"|'''PCR colo :''' |
- | + | - | |
+ | |bgcolor="#BCCDFD"|'''PCR colo :''' | ||
+ | - | ||
|- style="background: #d8d8d8; text-align: center;" | |- style="background: #d8d8d8; text-align: center;" | ||
- | |'''miniprep:''' | + | |bgcolor="#F8F8F8"|'''miniprep:''' |
- | + | STOPPED | |
- | |'''miniprep:''' | + | |bgcolor="#E0E3FE"|'''miniprep:''' |
- | + | - | |
- | |'''miniprep:''' | + | |bgcolor="#CBD1FD"|'''miniprep:''' |
- | + | - | |
+ | |bgcolor="#BCCDFD"|'''miniprep:''' | ||
+ | |||
+ | - | ||
|- style="background: #bebebe; text-align: center;" | |- style="background: #bebebe; text-align: center;" | ||
- | |'''sequencing :''' | + | |bgcolor="#F8F8F8"|'''sequencing :''' |
- | + | - | |
- | |'''sequencing :''' | + | |bgcolor="#E0E3FE"|'''sequencing :''' |
- | + | - | |
- | |'''sequencing :''' | + | |bgcolor="#CBD1FD"|'''sequencing :''' |
- | + | - | |
+ | |bgcolor="#BCCDFD"|'''sequencing :''' | ||
+ | |||
+ | - | ||
|} | |} | ||
+ | |||
Revision as of 18:40, 21 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
Time required : A lOT !!!!!
Experiments ran :
column 1 | column 2 | column 3 | column 4
|
PCR :
TatABCE matrix : | PCR : | PCR : | PCR : |
Verification on gel :
ok | Verification on gel :
- | Verification on gel :
- | Verification on gel :
- |
Purification on gel :
ok | Purification on gel :
- | Purification on gel :
- | Purification on gel :
-
|
digestion:
TatABCE | digestion: | digestion: | digestion: |
verification digestion:
ok | verification digestion:
- | verification digestion:
- | verification digestion:
-
|
ligation:
TatABCE | ligation: | ligation: | ligation: |
PCR colo :
ok | PCR colo :
- | PCR colo :
- | PCR colo :
- |
miniprep:
STOPPED | miniprep:
- | miniprep:
- | miniprep:
- |
sequencing :
- | sequencing :
- | sequencing :
- | sequencing :
- |
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
Time required : A lOT !!!!!
Experiments ran :
column 1 | column 2 | column 3 | column 4
|
PCR :
TatABCE matrix : | PCR : | PCR : | PCR : |
Verification on gel :
ok | Verification on gel :
- | Verification on gel :
- | Verification on gel :
- |
Purification on gel :
ok | Purification on gel :
- | Purification on gel :
- | Purification on gel :
-
|
digestion:
TatABCE | digestion: | digestion: | digestion: |
verification digestion:
ok | verification digestion:
- | verification digestion:
- | verification digestion:
-
|
ligation:
TatABCE | ligation: | ligation: | ligation: |
PCR colo :
ok | PCR colo :
- | PCR colo :
- | PCR colo :
- |
miniprep:
STOPPED | miniprep:
- | miniprep:
- | miniprep:
- |
sequencing :
- | sequencing :
- | sequencing :
- | sequencing :
- |
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
Time required : A lOT !!!!!
Experiments ran :
column 1 | column 2 | column 3 | column 4
|
PCR :
TatABCE matrix : | PCR : | PCR : | PCR : |
Verification on gel :
ok | Verification on gel :
- | Verification on gel :
- | Verification on gel :
- |
Purification on gel :
ok | Purification on gel :
- | Purification on gel :
- | Purification on gel :
-
|
digestion:
TatABCE | digestion: | digestion: | digestion: |
verification digestion:
ok | verification digestion:
- | verification digestion:
- | verification digestion:
-
|
ligation:
TatABCE | ligation: | ligation: | ligation: |
PCR colo :
ok | PCR colo :
- | PCR colo :
- | PCR colo :
- |
miniprep:
STOPPED | miniprep:
- | miniprep:
- | miniprep:
- |
sequencing :
- | sequencing :
- | sequencing :
- | sequencing :
- |