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EPF-Lausanne/9 September 2009
From 2009.igem.org
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(→Characterization) |
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RO1#1 + 0.5 TRP : | RO1#1 + 0.5 TRP : | ||
[[Image:RO1105TRP.jpg|center|RO1#1 +0.5 TRP]] | [[Image:RO1105TRP.jpg|center|RO1#1 +0.5 TRP]] | ||
| - | RO1#2 + 0.5 TRP shows the same tendancy. | + | RO1#2 + 0.5 TRP shows the same tendancy : |
| + | [[Image:RO1205TRP.jpg|center|RO1#2 +0.5 TRP]] | ||
RO1#1 + 1 TRP : | RO1#1 + 1 TRP : | ||
[[Image:RO111TRP.jpg|center|RO1#1 + 1 TRP]] | [[Image:RO111TRP.jpg|center|RO1#1 + 1 TRP]] | ||
| - | Again, RO1#2 clone has the same behaviour. | + | Again, RO1#2 clone has the same behaviour : |
| + | [[Image:RO121TRP.jpg|center|RO1#2 + 1 TRP]] | ||
RO1#1 without TRP: | RO1#1 without TRP: | ||
[[Image:RO11.jpg|center|RO1#1 - TRP]] | [[Image:RO11.jpg|center|RO1#1 - TRP]] | ||
| - | Clone #2 has the same behaviour. | + | Clone #2 has the same behaviour : |
| + | [[Image:RO12.jpg|center|RO1#2 - TRP]] | ||
RO1#3 clones doesn't follow these tendancy. It actually seems that this clone isn't working at all. It is possible a spontaneous mutation occured in some place... On all different conditions, RO1#3 clones show a flat curve, like RBS. It doesn't express RFP which means either our construct wasn't inserted, either there has been a mutation in it. For example + 1 TRP : | RO1#3 clones doesn't follow these tendancy. It actually seems that this clone isn't working at all. It is possible a spontaneous mutation occured in some place... On all different conditions, RO1#3 clones show a flat curve, like RBS. It doesn't express RFP which means either our construct wasn't inserted, either there has been a mutation in it. For example + 1 TRP : | ||
| Line 99: | Line 102: | ||
RO2#1 + 1.5 TRP : | RO2#1 + 1.5 TRP : | ||
[[Image:RO2115TRP.jpg|center|RO2#1 + 1.5 TRP]] | [[Image:RO2115TRP.jpg|center|RO2#1 + 1.5 TRP]] | ||
| + | RO2#3 in the same conditions : | ||
| + | [[Image:RO2315TRP.jpg|center|RO2#3 + 1.5 TRP]] | ||
RO2#1 + 0.5 TRP + 0.5 ATC : | RO2#1 + 0.5 TRP + 0.5 ATC : | ||
[[Image:RO2105TRP05ATC.jpg|center|RO2#1 + 0.5 TRP + 0.5 ATC]] | [[Image:RO2105TRP05ATC.jpg|center|RO2#1 + 0.5 TRP + 0.5 ATC]] | ||
| + | RO2#3 in the same conditions : | ||
| + | [[Image:RO2305TRP05ATC.jpg|center|RO2#3 + 0.5 TRP + 0.5 ATC]] | ||
RO2#1 + 0.5 TRP + 1 ATC : | RO2#1 + 0.5 TRP + 1 ATC : | ||
[[Image:RO2105TRP1ATC.jpg|center|RO2#1 + 0.5 TRP + 1 ATC]] | [[Image:RO2105TRP1ATC.jpg|center|RO2#1 + 0.5 TRP + 1 ATC]] | ||
| + | RO2#3 in the same conditions : | ||
| + | [[Image:RO2305TRP1ATC.jpg|center|RO2#3 + 0.5 TRP + 1 ATC]] | ||
RO2#1 + 1 TRP + 0.5 ATC : | RO2#1 + 1 TRP + 0.5 ATC : | ||
[[Image:RO211TRP05ATC.jpg|center|RO2#1 + 1 TRP + 0.5 ATC]] | [[Image:RO211TRP05ATC.jpg|center|RO2#1 + 1 TRP + 0.5 ATC]] | ||
| + | RO2#3 in the same conditions : | ||
| + | [[Image:RO231TRP05ATC.jpg|center|RO2#3 + 1 TRP + 0.5 ATC]] | ||
RO2#1 + 1 TRP + 1 ATC : | RO2#1 + 1 TRP + 1 ATC : | ||
[[Image:RO211TRP1ATC.jpg|center|RO2#1 + 1 TRP + 1 ATC]] | [[Image:RO211TRP1ATC.jpg|center|RO2#1 + 1 TRP + 1 ATC]] | ||
| + | RO2#3 in the same conditions : | ||
| + | [[Image:RO231TRP1ATC.jpg|center|RO2#3 + 1 TRP + 1 ATC]] | ||
RO2#1 + 0.5 ATC : | RO2#1 + 0.5 ATC : | ||
[[Image:RO2105ATC.jpg|center|RO2#1 + 0.5 ATC]] | [[Image:RO2105ATC.jpg|center|RO2#1 + 0.5 ATC]] | ||
| + | RO2#3 in the same conditions : | ||
| + | [[Image:RO2305ATC.jpg|center|RO2#3 + 0.5 ATC]] | ||
RO2#1 + 1 ATC : | RO2#1 + 1 ATC : | ||
[[Image:RO211ATC.jpg|center|RO2#1 + 1 ATC]] | [[Image:RO211ATC.jpg|center|RO2#1 + 1 ATC]] | ||
| + | RO2#3 in the same conditions : | ||
| + | [[Image:RO231ATC.jpg|center|RO2#3 + 1 ATC]] | ||
RO2#1 + 1.5 ATC : | RO2#1 + 1.5 ATC : | ||
[[Image:RO2115ATC.jpg|center|RO2#1 + 1.5 ATC]] | [[Image:RO2115ATC.jpg|center|RO2#1 + 1.5 ATC]] | ||
| + | RO2#3 in the same conditions : | ||
| + | [[Image:RO2315ATC.jpg|center|RO2#3 + 1.5 ATC]] | ||
Clone RO2#2 is not working, it has always a flat curve like this (for ex +0.5 ATC): | Clone RO2#2 is not working, it has always a flat curve like this (for ex +0.5 ATC): | ||
Revision as of 11:34, 2 October 2009
Wet Lab
Characterization
| With Atc | Without Atc | |||
|---|---|---|---|---|
| With Trp |
| 1/2 Trp 1 Trp 3/2 Trp | ||
| Without Trp | 1/2 Atc 1 Atc 3/2 Atc | Without Atc nor Trp |
Results of the characterization
For the RBS :
RBS was our negative control. We can see that there is no RFP fluorescence (because of course RBS has no RFP gene).
For LacI-RFP 1 + IPTG :
LacI-RFP 2 + IPTG show the same tendancy.
For Read Out 1
RO1#1 + 0.5 TRP :
RO1#2 + 0.5 TRP shows the same tendancy :
RO1#1 + 1 TRP :
Again, RO1#2 clone has the same behaviour :
RO1#1 without TRP:
Clone #2 has the same behaviour :
RO1#3 clones doesn't follow these tendancy. It actually seems that this clone isn't working at all. It is possible a spontaneous mutation occured in some place... On all different conditions, RO1#3 clones show a flat curve, like RBS. It doesn't express RFP which means either our construct wasn't inserted, either there has been a mutation in it. For example + 1 TRP :
For Read Out 2
RO2#1 without TRP :
RO2#3 has exactly the same curve shape :
RO2#1 + 0.5 TRP :
RO2#3 has a more "normal" shape :
RO2#1 + 1 TRP :
RO2#3 is the same :
RO2#1 + 1.5 TRP :
RO2#3 in the same conditions :
RO2#1 + 0.5 TRP + 0.5 ATC :
RO2#3 in the same conditions :
RO2#1 + 0.5 TRP + 1 ATC :
RO2#3 in the same conditions :
RO2#1 + 1 TRP + 0.5 ATC :
RO2#3 in the same conditions :
RO2#1 + 1 TRP + 1 ATC :
RO2#3 in the same conditions :
RO2#1 + 0.5 ATC :
RO2#3 in the same conditions :
RO2#1 + 1 ATC :
RO2#3 in the same conditions :
RO2#1 + 1.5 ATC :
RO2#3 in the same conditions :
Clone RO2#2 is not working, it has always a flat curve like this (for ex +0.5 ATC):
People in the lab
Mélanie, Caroline, Basile, Nicolas
