Difference between revisions of "Team:Glasgow/Practices"
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| − | <p class="links scrollOverview"><a style="color:blue;" href="https://2015.igem.org/Team:Glasgow"> Home</a> > <a style="color:blue;" href="https://2015.igem.org/Team:Glasgow/Project/Overview"> Project</a> > <a style="color:blue;" href="https://2015.igem.org/Team:Glasgow/Project/Overview/ | + | <p class="links scrollOverview"><a style="color:blue;" href="https://2015.igem.org/Team:Glasgow"> Home</a> > <a style="color:blue;" href="https://2015.igem.org/Team:Glasgow/Project/Overview"> Project</a> > <a style="color:blue;" href="https://2015.igem.org/Team:Glasgow/Project/Overview/Practices">Policy & Practices</a></p> |
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We used this opportunity to find out what our target market thought of Furri-Lux – to help with this, we had a large colouring-in picture of Furri-Lux’s logo for younger children, as well as blank paper for older kids to design their own ‘friendly monster’, all of which we displayed on a poster board next to our table. | We used this opportunity to find out what our target market thought of Furri-Lux – to help with this, we had a large colouring-in picture of Furri-Lux’s logo for younger children, as well as blank paper for older kids to design their own ‘friendly monster’, all of which we displayed on a poster board next to our table. | ||
These drawings later became inspiration for Furri-Lux’s outer design, as we were particularly interested in which features children associated with ‘friendly’ and ‘scary’ monsters. | These drawings later became inspiration for Furri-Lux’s outer design, as we were particularly interested in which features children associated with ‘friendly’ and ‘scary’ monsters. | ||
| − | For their parents, we had flyers with information about iGEM, the competition, past projects as well as our project to take away and read at their leisure, along with an interactive ‘circuit quiz’ on how the bioluminescence system worked inside the cells. The right answer had to be placed onto the board for it to conduct, and light up the LED! | + | <br/>For their parents, we had flyers with information about iGEM, the competition, past projects as well as our project to take away and read at their leisure, along with an interactive ‘circuit quiz’ on how the bioluminescence system worked inside the cells. The right answer had to be placed onto the board for it to conduct, and light up the LED! |
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| − | On the whole, many parents appeared very receptive to the idea of a children’s product incorporating bacteria, which was greatly encouraging. A common theme seemed to be that many parents believed their children were already exposed to a lot of much more harmful bacteria in every day life, so more education for younger kids was definitely needed. | + | On the whole, many parents appeared very receptive to the idea of a children’s product incorporating bacteria, which was greatly encouraging. A common theme seemed to be that many parents believed their children were already exposed to a lot of much more harmful bacteria in every day life, so more education for younger kids was definitely needed.<br/> |
| − | **Should I keep this next part?** However, their concern was apparent when we tried to talk to the children about ‘monsters under the bed’ – clearly this was a fear that parents were not immediately willing to validate, and they had put a lot of effort into trying to convince their children that these ‘monsters’ didn’t exist. Therefore, buying a product that offered help with this problem seemed contradictory and counter-productive. We at iGEM Glasgow, on the other hand, feel that validating the children’s’ fears is more important to their healthy emotional development, as well as strengthening the parent-child bond as they work through it together. | + | <strong>**Should I keep this next part?**</strong> However, their concern was apparent when we tried to talk to the children about ‘monsters under the bed’ – clearly this was a fear that parents were not immediately willing to validate, and they had put a lot of effort into trying to convince their children that these ‘monsters’ didn’t exist. Therefore, buying a product that offered help with this problem seemed contradictory and counter-productive. <br/>We at iGEM Glasgow, on the other hand, feel that validating the children’s’ fears is more important to their healthy emotional development, as well as strengthening the parent-child bond as they work through it together. |
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Parents also expressed concerns that curious children would try to open the container themselves – which led us to design the removable handle for the bacteria’s release. To read more about this, see our <a href = “2015.igem.org/Team:Glasgow/Design”>Product Design</a> page. | Parents also expressed concerns that curious children would try to open the container themselves – which led us to design the removable handle for the bacteria’s release. To read more about this, see our <a href = “2015.igem.org/Team:Glasgow/Design”>Product Design</a> page. | ||
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| − | <h2> | + | <h2>Hyndland School Visit</h2> |
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| − | + | We had the chance to invite some senior pupils from the nearby Hyndland Secondary School to our lab, so that they could learn more about the iGEM competition and the other opportunities available to them at university.<br/><br/> | |
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| − | + | Afterwards, we let them practice their aseptic technique by drawing their own pictures on agar plates with our glowing bacteria – and they came up a treat!<br/><br/> | |
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| + | Most of the pupils were studying a course on Lab Skills, so we showed them around the lab we were working in and discussed the operation of some of the equipment they may not have had substantial access to in school – such as the autoclave and flow hood.<br/><br/> | ||
| + | The remaining pupils were from their Advanced Biology course, who found the more in depth presentation of our bioluminescence system to be more interesting and insightful. | ||
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| − | <h2> | + | <h2> Survey Analysis</h2> |
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Revision as of 12:42, 18 September 2015
Home > Project > Policy & Practices
Glasgow Science Centre
On the 1st and 2nd of August, we presented a stall at Glasgow Science Centre as part of their ‘Meet the Experts’ event. This was our first chance to reach out and engage with the public to get real feedback about the things we had planned, which was overwhelmingly positive!
We used this opportunity to find out what our target market thought of Furri-Lux – to help with this, we had a large colouring-in picture of Furri-Lux’s logo for younger children, as well as blank paper for older kids to design their own ‘friendly monster’, all of which we displayed on a poster board next to our table.
These drawings later became inspiration for Furri-Lux’s outer design, as we were particularly interested in which features children associated with ‘friendly’ and ‘scary’ monsters.
For their parents, we had flyers with information about iGEM, the competition, past projects as well as our project to take away and read at their leisure, along with an interactive ‘circuit quiz’ on how the bioluminescence system worked inside the cells. The right answer had to be placed onto the board for it to conduct, and light up the LED!
On the whole, many parents appeared very receptive to the idea of a children’s product incorporating bacteria, which was greatly encouraging. A common theme seemed to be that many parents believed their children were already exposed to a lot of much more harmful bacteria in every day life, so more education for younger kids was definitely needed.
**Should I keep this next part?** However, their concern was apparent when we tried to talk to the children about ‘monsters under the bed’ – clearly this was a fear that parents were not immediately willing to validate, and they had put a lot of effort into trying to convince their children that these ‘monsters’ didn’t exist. Therefore, buying a product that offered help with this problem seemed contradictory and counter-productive.
We at iGEM Glasgow, on the other hand, feel that validating the children’s’ fears is more important to their healthy emotional development, as well as strengthening the parent-child bond as they work through it together.
Parents also expressed concerns that curious children would try to open the container themselves – which led us to design the removable handle for the bacteria’s release. To read more about this, see our Product Design page.
Hyndland School Visit
We had the chance to invite some senior pupils from the nearby Hyndland Secondary School to our lab, so that they could learn more about the iGEM competition and the other opportunities available to them at university.
Afterwards, we let them practice their aseptic technique by drawing their own pictures on agar plates with our glowing bacteria – and they came up a treat!
Most of the pupils were studying a course on Lab Skills, so we showed them around the lab we were working in and discussed the operation of some of the equipment they may not have had substantial access to in school – such as the autoclave and flow hood.
The remaining pupils were from their Advanced Biology course, who found the more in depth presentation of our bioluminescence system to be more interesting and insightful.
Survey Analysis
Originally, the system containing UirS, UirR, and PlsiR accounts for a negative phototactic response to unidirectional UV-A light. The proposed mechanism puts UirS, a transmembrane protein of the CBCR family, as the molecule that perceives UV light. It is suggested that through a physical interaction between UirS and UirR and possibly a phosphotransfer from UirS to UirR, UirR is released from the transmembrane protein. The released UirR can now bind to DNA and UirR, which is similar to other activators of stress responses, was found to be a transcriptional activator of lsiR after binding to its promoter PlsiR .
We suggest a system where UV-light triggers the expression of a repressor that acts on the production of bioluminescence genes to alleviate the burden they may cause on the cells’ metabolism if constantly expressed. During the night, the bioluminescence genes are expressed, and produce a green-blue light. As it becomes day, UV-A causes the release and activation of UirR. UirR binds to PlsiR to turn expression of the repressor PhlF. PhlF binds to PPhlF to turn off expression of LuxCDABE, so there is no bioluminescence. As it becomes night again, UirR is no longer bound to PlsiR so expression of PhlF is turned off. As PPhlF is no longer repressed, expression of LuxCDABE is turned and bioluminescence is produced again.
We have confirmed through the use of a laser scanner that PlsiR is not active when UirS and UirR are absent. PlsiR was ligated to GFP with two ribosome binding sites of different strength and no fluorescence was observed (the parts we used for this experiment were K1725401 and K1725402) (Chart 1). Moreover, cells that possess UirR but lack UirS also did not show levels of fluorescence above the expected for E. coli. Therefore, UirR is not sufficient to drive the activation of PlsiR.
Survivability
Introduction
The fact that UV exposure can be lethal to E.coli is well documented. After deciding to use a UV sensor to activate our system, it became obvious that we would have to examine the effects of exposing E.coli to the amount of UV needed to activate the sensor over time. To do this we generated a number of survival curves. (Please note, these experiments were not repeated enough to generate statistically significant results.) The graphs have been included as an indication of the thought process that went into guiding the experiments. Given more time we would have repeated these experiments many more times. Genotypes of the strains used can be found here for MG6115 TOP10 and DH5α and here for DS941.
Initial aims
Initially we wanted to explore what happened over a relatively small period of exposure, as our assumption was that we would see substantial reduction in a fixed number of E.coli, even over a relatively small time period. We also wanted to see how the different strains (DH5α ,TOP10) being used would respond compared to each other.
Since both DH5α and TOP10 are recA negative, and are therefore incapable of certain major kinds of DNA repair, we predicted that these strains would be more acutely affected than DS941 and MG6155.
Method
1ml 10x serial dilutions were made up to 10-6 from a 5ml overnight of each strain. 10µl spots of each dilution were then spotted onto LB agar plates.
These plates were then exposed to 50µmoles/m2/s of UVA at room temperature in illumination cabinets. Time points were then taken by removing plates from the illumination cabinet.
After illumination, plates were incubated at 37°C, under the assumption that every single viable cell will form a colony. The length of incubation is irrelevant, provided that every cell is given enough time to form a visible colony. This also forms the basis of our counting system, where a colony is assumed to have come from a single cell. We also make the assumption that cell division at the temperature and time we were running the experiment was negligible/nonexistent.
After incubation, the colonies on each spot/dilution were counted. The number of colonies from the lowest visible dilution (some dilutions formed a lawn of growth) were then multiplied by the dilution factor to approximate how many cells would be in 10µl of undiluted culture.
Results
Conclusion
Read More!
