McGill/Results
From 2007.igem.org
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This year our new team has been developing the project established last year which you can see on the McGill 2006 wiki. We have made many new discoveries and have run into some problems hindering our progress along the way; nevertheless, we were able to make some new developments as outlined below: | This year our new team has been developing the project established last year which you can see on the McGill 2006 wiki. We have made many new discoveries and have run into some problems hindering our progress along the way; nevertheless, we were able to make some new developments as outlined below: | ||
| - | 1. Reproduction of Last Year's System with a now known caveat. <br> | + | '''1. Reproduction of Last Year's System with a now known caveat. <br>''' |
The primary oscillating system, the I15004 with the J40001 as outlined in the previous pages was produced last year and viewed using a microscope for several hours to analyze the oscillatory period and confirm the presence of these oscillations. This year, we took this a step further. Using a spectrophotometer, we were able to achieve more precise quantitative results for the oscillating network and we were also able to test many samples at a time with variations in the testing conditions: variable concentrations of cells, variable concentrations of IPTG, Dox (tetracycline analog), and AHL (synthetic auto-inducer). From these experiments, we were able to come up with some interesting results.<br> | The primary oscillating system, the I15004 with the J40001 as outlined in the previous pages was produced last year and viewed using a microscope for several hours to analyze the oscillatory period and confirm the presence of these oscillations. This year, we took this a step further. Using a spectrophotometer, we were able to achieve more precise quantitative results for the oscillating network and we were also able to test many samples at a time with variations in the testing conditions: variable concentrations of cells, variable concentrations of IPTG, Dox (tetracycline analog), and AHL (synthetic auto-inducer). From these experiments, we were able to come up with some interesting results.<br> | ||
#The more concentrated the cells, the worse the apparent oscillations.<br> | #The more concentrated the cells, the worse the apparent oscillations.<br> | ||
#The presence of Dox seemed to stabilize the oscillations. <br> | #The presence of Dox seemed to stabilize the oscillations. <br> | ||
#The greater the amount of AHL, the worse the apparent oscillations as predicted with the simulations. <br> | #The greater the amount of AHL, the worse the apparent oscillations as predicted with the simulations. <br> | ||
| - | # | + | #Using an optical density assay, it was found that the cells were in fact dying (or clumping) thus lowering the optical density through the testing period. <br> |
| - | 2. Production of a new I15004 brick to compensate for the mal-functioning biobrick. <br> | + | #Oscillations occurs also without the presence of the I15004 brick (thus, the J40001 brick oscillates on its own. The reason for this is attributed to the expression of the endogenous lac in the BL21 cells, the expression cells used in this experiment. So, to remedy this, we replaced the BL21 cells with MC cells, the lac-, tet- cells used by Elowitz himself when doing these experiments. Results: no oscillations which led us to the nest point, the malfunctioning I15004 brick needs to be replaced. |
| - | 3. Synthesis of a new Represillator from different components parts, again, to compensate for the mal-function biobrick. <br> | + | '''2. Production of a new I15004 brick to compensate for the mal-functioning biobrick. <br>''' |
| - | 4. Synthesis and completion of a new biobrick, an addition to the J40001 made last years with the addition of Aiia.<br> | + | '''3. Synthesis of a new Represillator from different components parts, again, to compensate for the mal-function biobrick. <br>''' |
| + | '''4. Synthesis and completion of a new biobrick, an addition to the J40001 made last years with the addition of Aiia.<br>''' | ||
Revision as of 16:14, 26 October 2007
This year our new team has been developing the project established last year which you can see on the McGill 2006 wiki. We have made many new discoveries and have run into some problems hindering our progress along the way; nevertheless, we were able to make some new developments as outlined below:
1. Reproduction of Last Year's System with a now known caveat.
The primary oscillating system, the I15004 with the J40001 as outlined in the previous pages was produced last year and viewed using a microscope for several hours to analyze the oscillatory period and confirm the presence of these oscillations. This year, we took this a step further. Using a spectrophotometer, we were able to achieve more precise quantitative results for the oscillating network and we were also able to test many samples at a time with variations in the testing conditions: variable concentrations of cells, variable concentrations of IPTG, Dox (tetracycline analog), and AHL (synthetic auto-inducer). From these experiments, we were able to come up with some interesting results.
- The more concentrated the cells, the worse the apparent oscillations.
- The presence of Dox seemed to stabilize the oscillations.
- The greater the amount of AHL, the worse the apparent oscillations as predicted with the simulations.
- Using an optical density assay, it was found that the cells were in fact dying (or clumping) thus lowering the optical density through the testing period.
- Oscillations occurs also without the presence of the I15004 brick (thus, the J40001 brick oscillates on its own. The reason for this is attributed to the expression of the endogenous lac in the BL21 cells, the expression cells used in this experiment. So, to remedy this, we replaced the BL21 cells with MC cells, the lac-, tet- cells used by Elowitz himself when doing these experiments. Results: no oscillations which led us to the nest point, the malfunctioning I15004 brick needs to be replaced.
2. Production of a new I15004 brick to compensate for the mal-functioning biobrick.
3. Synthesis of a new Represillator from different components parts, again, to compensate for the mal-function biobrick.
4. Synthesis and completion of a new biobrick, an addition to the J40001 made last years with the addition of Aiia.
