IGEM:Imperial/Results/S01656: Difference between revisions
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===[[IGEM:IMPERIAL/Protocols/S01656|S01656 Protocol Page]]=== | ===[[IGEM:IMPERIAL/Protocols/S01656|S01656 Protocol Page]]=== | ||
==Sunday, 20 August 2006== | ==Sunday, 20 August 2006== | ||
| Line 74: | Line 66: | ||
::% Difference =<font color=green> 4.25%</font> | ::% Difference =<font color=green> 4.25%</font> | ||
::Conclusion: <font color=red>Not statistically significant</font> | ::Conclusion: <font color=red>Not statistically significant</font> | ||
::*'''[[User:TomH|Tom]] 13:58, 26 August 2006 (EDT)''': [[IGEM:IMPERIAL/2006/project/Oscillator/Problems/aiiA|Debateable]] | |||
*Resolution | *Resolution | ||
| Line 79: | Line 72: | ||
:Test expression of AiiA protein by SDS electrophoresis | :Test expression of AiiA protein by SDS electrophoresis | ||
:Re-test the protocol on Wednesday 23 August 2006 with higher concentration of AHL | :Re-test the protocol on Wednesday 23 August 2006 with higher concentration of AHL | ||
==Wednesday, 23 August 2006== | |||
===Results of western Blot=== | |||
[[Image:Non-working Gel.JPG]] | |||
The IPTG has '''not''' induced the production of a protein which weighs 28736 kd ''(aiiA)'' this should be loacted inside the oval. We are repeating the expperiment to confirm the results. We are sequencing the part to confirm the results. The ladder is invitrogen marker 12. | |||
[[User:JohnChattaway|JohnChattaway]] 10:13, 23 August 2006 (EDT) | |||
===Results of AHL Testing=== | |||
*Used 100 {{uM}} stock concentration instead of 10 {{uM}}. Used 800 {{uL}} of supernatant and 1200 {{uL}} of T9002 cells. | |||
* Normalised Fluorescence of +IPTG +AHL | |||
::Tube 1 - 4353 | |||
::Tube 2 - 3552 | |||
::Tube 3 - 4476 | |||
*Normalised Fluorescence of +IPTG -AHL | |||
::Tube 1 - 4157 | |||
*Normalised Fluorescence of -IPTG +AHL | |||
::Tube 1 - 3688 | |||
::Tube 2 - 4609 | |||
::Tube 3 - 4197 | |||
*Normalised Fluroescence of -IPTG -AHL | |||
::Tube 1 - 3608 | |||
*Comparison between +IPTG +AHL and -IPTG +AHL: | |||
::% Difference =<font color=green> 0.91%</font> | |||
::Conclusion: <font color=red>Not statistically significant</font> | |||
*Resolution | |||
:Re-test the protocol on Thursday 24 August 2006 with same concentration of AHL | |||
Latest revision as of 10:58, 26 August 2006
S01656 Protocol Page
Sunday, 20 August 2006
- Normalised Fluorescence of +IPTG +AHL
- Tube 1 - 233
- Tube 2 - 237
- Tube 3 - 239
- Normalised Fluorescence of +IPTG -AHL
- Tube 1 - 219
- Normalised Fluorescence of -IPTG +AHL
- Tube 1 - 244
- Normalised Fluroescence of -IPTG -AHL
- Tube 1 - 218
- Comparison between +IPTG +AHL and -IPTG +AHL:
- % Difference = 2.94%
- Conclusion: Not statistically significant
- Resolution
- Test expression of AiiA protein by SDS electrophoresis
- Re-test the protocol on Tuesday 22 August 2006
Tuesday, 22 August 2006
- Different formulation used to calculate normalised fluorescence. Used the same calculations as the T9002. See updated spreadsheet.
- Normalised Fluorescence of +IPTG +AHL
- Tube 1 - 4409
- Tube 2 - 4659
- Tube 3 - 4574
- Normalised Fluorescence of +IPTG -AHL
- Tube 1 - 4574
- Normalised Fluorescence of -IPTG +AHL
- Tube 1 - 4769
- Tube 2 - 4762
- Tube 3 - 4834
- Normalised Fluroescence of -IPTG -AHL
- Tube 1 - 4445
- Comparison between +IPTG +AHL and -IPTG +AHL:
- % Difference = 4.25%
- Conclusion: Not statistically significant
- Tom 13:58, 26 August 2006 (EDT): Debateable
- Resolution
- Test expression of AiiA protein by SDS electrophoresis
- Re-test the protocol on Wednesday 23 August 2006 with higher concentration of AHL
Wednesday, 23 August 2006
Results of western Blot
The IPTG has not induced the production of a protein which weighs 28736 kd (aiiA) this should be loacted inside the oval. We are repeating the expperiment to confirm the results. We are sequencing the part to confirm the results. The ladder is invitrogen marker 12.
JohnChattaway 10:13, 23 August 2006 (EDT)
Results of AHL Testing
- Used 100 μM stock concentration instead of 10 μM. Used 800 μL of supernatant and 1200 μL of T9002 cells.
- Normalised Fluorescence of +IPTG +AHL
- Tube 1 - 4353
- Tube 2 - 3552
- Tube 3 - 4476
- Normalised Fluorescence of +IPTG -AHL
- Tube 1 - 4157
- Normalised Fluorescence of -IPTG +AHL
- Tube 1 - 3688
- Tube 2 - 4609
- Tube 3 - 4197
- Normalised Fluroescence of -IPTG -AHL
- Tube 1 - 3608
- Comparison between +IPTG +AHL and -IPTG +AHL:
- % Difference = 0.91%
- Conclusion: Not statistically significant
- Resolution
- Re-test the protocol on Thursday 24 August 2006 with same concentration of AHL
