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Team:Calgary/Project/Collaboration
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Based on the results from this experiment and from our previous attempt, we're confident that this method has potential. Ampicillin-resistance was used as a selection marker, the plasmids containing RFP also contained a gene for Amp-resistance. The lack of red colonies does not necessarily indicate a lack of transformation, since cells were still able to grow, chances are that they did not express the somewhat toxic RFP. </p> | Based on the results from this experiment and from our previous attempt, we're confident that this method has potential. Ampicillin-resistance was used as a selection marker, the plasmids containing RFP also contained a gene for Amp-resistance. The lack of red colonies does not necessarily indicate a lack of transformation, since cells were still able to grow, chances are that they did not express the somewhat toxic RFP. </p> | ||
| - | <img src= "https://static.igem.org/mediawiki/2014/2/20/PLATES1234.png"> | + | <img src= "https://static.igem.org/mediawiki/2014/2/20/PLATES1234.png" class= "Center"> |
<p>We believe this protocol is cost effective, the calcium citrate tablets cost us approximately $0.21. We submitted our data to the University of Columbia ‘s iGEM team and received a collaboration sticker. The winner will be announced in Boston, we can’t wait to see what other teams came up with!</p> | <p>We believe this protocol is cost effective, the calcium citrate tablets cost us approximately $0.21. We submitted our data to the University of Columbia ‘s iGEM team and received a collaboration sticker. The winner will be announced in Boston, we can’t wait to see what other teams came up with!</p> | ||
Revision as of 22:58, 17 October 2014
Collaboration
University of Virginia’s iGEM team: Collaborative Survey
The University of Virginia’s 2014 iGEM team designed a short online survey in order to learn about prevailing attitudes toward synthetic biology in the general public. Through this survey, they aimed to help future teams develop targeted educational events regarding synthetic biology.
We(the iGEM Calgary team) helped out by surveying over 40 people during our outreach events at the Telus Sparks Centre! Many people we encountered had misconceptions about synthetic biology. We were able to explain to them the significance of synthetic biology in medicine, the food industry and the environment. We are looking forward to hearing about the results of the survey and the University of Virginia’s iGEM teams suggestions for future educational events!
University of Cornell: Humans and Synbio
The University of Cornell created the”Humans and Synbio” facebook page to learn about peoples opinions regarding different aspects of synthetic biology. They asked iGEM teams from all around the world to participate!
We (iGEM Calgary) helped out by asking Calgarians their favorite application of synthetic biology, containment concerns and how they feel about ethical issues surrounding genetically engineered organisms. A few of the responses we got are posted below. To view more pictures please visit the “ Humans and Synbio” facebook page!
Do you think there is a limit in how far we should go with syn bio?
"Synthetic biology is an area of research which has unlimited potential for growth and innovation. I believe that synthetic biology has the capacity to be hugely advantageous to the scientific community and to society as a whole. It would be irresponsible to ignore the potentially harmful applications of synthetic biology, however, the potential benefits provided by research in this area exceed the disadvantages. Through establishing firm regulations in this area I believe that research can be conducted in a safe and productive manner. I believe it is the responsibility of members of society, including those in the scientific community, to establish the ethical groundwork which will shape the aims and applications of synthetic biology."
In general, how do you feel about humans purposely modifying organisms or species so they will be more useful to us?
"If humans can benefit from modifying organisms or species they should, as long as it is done in a humane sustainable way."
How would you define humane use of animals?
“By humane I mean in a way that doesn't cause unnecessary suffering or excruciating pain to the animals because life is precious and shouldn't be mistreated if we can help it.”
What is you favorite application of synthetic biology?
“I know it's an old school example, but my favorite application of syn bio would have to be when they got E. coli to make human insulin. That definitely revolutionized how we look at type 1 diabetes and really improved people's lives.”
Synthetic biology creates biological systems that do not naturally exist. Does anything about synthetic biology make you feel uncomfortable?
“The idea of synthetic biology doesn’t make me feel uncomfortable but the applications and potential outcomes are somewhat worrisome. I don’t know much about the topic on hand but just as long as there are regulations in place and governing bodies to overlook the research and applications, I would feel at ease with it all.”
How do you feel about humans purposely modifying organisms or species so that they will be more useful to us?
"I’m sure that there are benefits that do go along with that type of research, but I am a firm believer in the earth is where it is for a reason and we are where we are for a reason and life has evolved over millions of years without us having to make it better. We’ve made a lot of mistakes along the way so I don’t know if I trust research and peoples’ judgment. (I explained to her that domesticating animals is a way we’ve done this in the past, to which she responded…) I think all that would’ve naturally fallen into place anyways. Dogs really do love humans and they need to connect with a pack. Horses, if they really didn’t want to be trained, I don’t know if you could train them. Whereas, I believe they feel good when they’re serving a purpose as well. So I think those connections would’ve happened automatically. I don’t know if we can take complete credit for those modifications. I guess what I’m thinking is more that horses were the size of a dog and they evolved into the size they are today. I don’t know. I’m all for research certainly, but there’s also the question how far will it go. Is that going to be to evolve to the perfect human? And whose perception is the perfect human?"”
University of Columbia: Low budget iGEM Contest
The University of Columbia’s team challenged iGEM teams to figure out a cost effective way to transform bacteria, extract DNA and amplify genes of interest in order to make molecular biology globally available. Of course team Calgary accepted the challenge! We attempted to make competent cells using calcium supplements available at the local pharmacy.
Attempt #1: There were many types and blends of calcium supplements at the pharmacy. Our first choice was calcium-carbonate. We soon discovered Calcium carbonate has very low solubility which led to unsuccessful transformation.
Attempt #2: Our second choice was to use calcium citrate which is physiologically compatible and has a relatively high solubility. Physiological compatibility was important because we didn't want anything that could potentially harm our bacteria. Unfortunately, the pills contained a lot of filler, therefore we had to crush several pills before we got the desired concentration of calcium. The calcium solution made using the pills was cloudy and contained a lot of debris. Transformation with this solution was successfully but had very low efficiency. We tried again, but this time we centrifuged the solution in order to obtain a clear solution.
We performed our experiment with 4 different concentrations of calcium from the calcium citrate tablets (5mM, 10mM, 50mM, 100mM). For our positive control we used 50mM calcium chloride. We followed the protocol we use in our lab to make competent cells, this protocol is available under “general protocols.”
Below are three photos of three different treatments of E. coli to produce competent cells. We tested competency by transforming with RFP (BBa_J04450), which yields red cells. The positive control/standard (50mM calcium chloride) showed efficient transformation, which was replicated in the 50mM calcium-citrate treatment. For comparison, the 10mM calcium-citrate treatment yielded only a few colonies, though they were not red. Based on the results from this experiment and from our previous attempt, we're confident that this method has potential. Ampicillin-resistance was used as a selection marker, the plasmids containing RFP also contained a gene for Amp-resistance. The lack of red colonies does not necessarily indicate a lack of transformation, since cells were still able to grow, chances are that they did not express the somewhat toxic RFP.
We believe this protocol is cost effective, the calcium citrate tablets cost us approximately $0.21. We submitted our data to the University of Columbia ‘s iGEM team and received a collaboration sticker. The winner will be announced in Boston, we can’t wait to see what other teams came up with!
