Team:Cambridge/HumanPractices/MarketResearch

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As part of our market research we interviewed Dr. Konrad Siegfried from the ARSOlux Team. Check out the interview [[http://2012.igem.org/Team:Cambridge/HumanPractices/Interview HERE!]]
Below is a summary of our research into the suitability of our system for use in the field.  
Below is a summary of our research into the suitability of our system for use in the field.  

Revision as of 03:36, 27 October 2012

Parts for a reliable and field ready biosensing platform

Implementation of biosensors in real world situations has been made difficult by the unpredictable and non-quantified outputs of existing solutions, as well as a lack of appropriate storage, distribution and utilization systems. This leaves a large gap between a simple, functional sensing mechanism and a fully realised product that can be used in the field. We aim to bridge this gap at all points by developing a standardised ratiometric luciferase output in a Bacillus chassis. This output can be linked up with prototyped instrumentation and software for obtaining reliable quantified results. Additionally, we have reduced the specialized requirements for the storage and distribution of our bacteria by using Bacillus' sporulation system. To improve the performance of our biosensing platform we have genetically modified Bacillus’ germination speed. Lastly, we demonstrated the robustness of our system by testing it with a new fluoride riboswitch, providing the opportunity to tackle real life problems.

One minute tour! :)

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Contents

Judging Form

  • Please help the judges by filling out this form. Tell them what medal you think you deserve and why. Tell them which special prizes you should win. Help them find your best parts. Show them how you thought about the safety of your project. Helping the judges will help you too.

  • Team: Cambridge
  • Region: Europe
  • iGEM Year:2012
  • Track:Foundational Advance
  • Project Name:Parts for a reliable and field ready biosensing platform
  • Project Abstract: Implementation of biosensors in real world situations has been made difficult by the unpredictable and non-quantified outputs of existing solutions, as well as a lack of appropriate storage, distribution and utilization systems. This leaves a large gap between a simple, functional sensing mechanism and a fully realised product that can be used in the field.

    We aim to bridge this gap at all points by developing a standardised ratiometric luciferase output in a Bacillus chassis. This output can be linked up with prototyped instrumentation and software for obtaining reliable quantified results. Additionally, we have reduced the specialized requirements for the storage and distribution of our bacteria by using Bacillus' sporulation system. To improve the performance of our biosensing platform we have genetically modified Bacillus’ germination speed. Lastly, we demonstrated the robustness of our system by testing it with a new fluoride riboswitch, providing the opportunity to tackle real life problems.

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iGEM Medals for non-software teams

  • We believe our team deserves the following medal:
    • Bronze
    • Silver
    • √Gold

Because we met the following criteria (check all that apply and provide details where needed)

Requirements for a Bronze Medal

  • √Register the team, have a great summer, and plan to have fun at the Regional Jamboree.
  • √Successfully complete and submit this iGEM 2012 Judging form.
  • √Create and share a Description of the team's project using the iGEM wiki and the team's parts using the Registry of Standard Biological Parts.
  • √Plan to present a Poster and Talk at the iGEM Jamboree.
  • √Enter information detailing at least one new standard BioBrick Part or Device in the Registry of Standard Biological Parts. Including:
    • √Primary nucleaic acid sequence
    • √Description of function
    • √Authorship
    • Safety notes, if relevant.
    • √Acknowedgment of sources and references
  • √Submit DNA for at least one new BioBrick Part or Device to the Registry.

Additional Requirements for a Silver Medal

  • √Demonstrate that at least one new BioBrick Part or Device of your own design and construction works as expected; characterize the operation of your new part/device.
  • √Enter this information and other documentation on the part's 'Main Page' section of the Registry
    Part Number(s): BBa_K911004

Additional Requirements for a Gold Medal: (one OR more)

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iGEM Prizes

All teams are eligible for special prizes at the Jamborees. more... To help the judges, please indicate if you feel you should be evaluated for any of the following special prizes:

  • √Best Human Practice Advance
  • √Best Experimental Measurement
  • Best Model

Please explain briefly why you should receive any of these special prizes:

Best Human Practice Advance:

We feel that we deserve this prize for three reasons:

  1. We explored the impacts, *both positive and negative*, of synthetic biology as a solution to real world problems, through interviewing professionals working in a relevant field, namely the impact of arsenic water contamination in Bangladesh.
  2. We recognized existing problems with the way the current direction of synthetic. On going through the registry we found that most of the characterization data for biosensing parts is often neither comparable nor replicable. We have worked to solve this issue, for example with our ratiometric dual channel output.
  3. *Our project doesn’t stop here*, in Chanel number 6 (Team:Cambridge/HumanPractices/FutureDirections) we considered the future implications and technological applications of our project, as well as the means by which it could be improved by subsequent users. We feel that the end to an iGEM project should not be the conclusion of an idea, but the start of it.

Best BioBrick Measurement Approach:

It is absolutely vital that a quantitative, numerical, robust, and flexible measurement approach exists to relay information to a user that is an accurate representation of the input processed by a biological device. Working from these principles, the following was done:

  1. We designed and built Biologger, a *cheap, arduino-based, fully functional automatic rotary device* that has an incorporated ratiolumnometer
  2. Our project is entirely open-sourced and open-platform. We have published source code for the two applications which serve to operate the device, one for PCs and the other for Android devices, as well as the open source circuit design that provides this ratiometric reading. Furthermore, the Android app is able to receive its data wirelessly, which we feel is a great advance in BioBrick measurement.
  3. Our dual-channel luciferase reporter was successfully tested with a dilution series of E.coli transformed with the Lux Operon (under pBAD) biobrick (Part BBa_K325909) of the Cambridge iGEM 2010 team. It can detect, with good accuracy, both different light intensities, as well as the percentages of blue or orange frequencies in a sample.
  4. Our device was successfully tested using artificial light to detect different frequencies (colours) as well.

Having done all the above, we believe that this fully open-sourced instrumentation kit (mechanical) chassis, electronics, software code), estimated at *$35.00* (or $85.00 if a Bluetooth modem is required), is a complete BioBrick measurement solution for any and all BioBricks with a light output.

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Team_Parts

To help the judges evaluate your parts, please identify 3 of your parts that you feel are best documented and are of the highest quality.

  • Best new BioBrick part (natural)
    BBa_K911003
    Best new BioBrick part (engineered)
    BBa_K911004
  • Best improved part(s): None

List any other parts you would like the judges to examine:BBa_K911001, BBa_K911009, BBa_K911008

Please explain briefly why the judges should examine these other parts:

  • Magnesium Sensitive Riboswitch BBa_K911001
    As a riboswitch sensing construct, this part is an entirely new type of biosensor (along with the fluoride construct) that could potentially change the way we think about designing input genetic circuits. Unlike the fluoride riboswitch, it is a derepression system and therefore serves to demonstrate the principle that riboswitches can be used regardless of whether they turn on or off their reporter.
  • Fluorescent ratiometric construct for standardizing promoter output BBa_K911009
    Fluorescence is a major cornerstone for biosensors in the registry, however, most parts do not involve the use of a ratiometric output, which has been shown in the literature to provide much more reliable and meaningful data. This part not only furthers the development of ratiometric measurements in molecular biology but due to the choice of promoters and terminators it can be used to characterize the difference in activity between E. coli and B. Subtilis
  • Fast Germination (B. subtilis) BBa_K911008
    This part is entirely novel for the registry and fully utilizes the recombination machinery inherent in the Bacillus chassis. Have spores that can germinate at a faster rate is certainly a worthy achievement and could help with experiments with B. Subtilis that any future iGEM teams may wish to perform.

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iGEM Safety

For iGEM 2012 teams are asked to detail how they approached any issues of biological safety associated with their projects.

The iGEM judges expect that you have answered the four safety questions Safety page on your iGEM 2012 wiki.

Please provide the link to that page: Page name: Team:Cambridge/Safety

Attribution and Contributions

For iGEM 2012 the description of each project must clearly attribute work done by the team and distinguish it from work done by others, including the host labs, advisors, and instructors.

Please provide the link to that page, or comments in the box below: Page name: Team:Cambridge/Attributions

Comments

If there is any other information about your project you would like to highlight for the judges, please provide a link to your wiki page here: Team:Cambridge/Overview/DesignProcess

Human Practices – Groundwater contamination in rural India

As part of our market research we interviewed Dr. Konrad Siegfried from the ARSOlux Team. Check out the interview [HERE!]

Below is a summary of our research into the suitability of our system for use in the field.

Finding a Market

The potential use of our system in this context first came to light when we became aware of WaterLifeIndia Ltd who won an award for inclusive business models at the G20 conference earlier this year for their work in providing clean, safe water in response to groundwater contamination for B.O.P. (economic base of the pyramid) customers in India.

We then contacted the British Red Cross, often the first on the ground when disaster strikes, to ask about the human impact of water contamination. When first arriving, all water must be treated as contaminated until it has been tested and shown not to be. For people living in areas with chronic contamination problems, finding clean water is an incredibly time consuming process which can take many hours out of peoples (predominantly women’s) days.

Further research showed that the problem is extensive and as of April 2011, Arsenic, Fluoride, Iron, Salinity and Nitrate groundwater contamination continued to be a problem in many states – especially in rural areas. Before these problems can be tackled, the scope of the problem must first be determined, requiring sensitive detection apparatus.

The consequences of fluoride contamination of ground water. Fluoride has been shown to damage the enamel of teeth in children between the ages of 0-8. This dental fluorosis is often accompanied by skeletal fluorosis, associated with skeletal pain and problems with movement. Excessive fluoride ingestion has also been linked to neurological damage in young children.


Who is our market?

Groundwater contamination can be caused by a variety of factors ranging from natural disasters to improper disposal of industrial waste. Within the topic of groundwater contamination therefore, there is still substantial variety among potential customers. We anticipate that these customers will be looking at levels of a variety of contaminants in different areas though we anticipate that they will need their testing system to provide reliable, accurate quantitative data about more than one contaminant and that testing will probably involve multiple tests over a prolonged time span. Customers may include researchers, charity workers, public health workers or heavy industries aiming to reduce their environmental impact.


What are the current options available?

Because of the rural nature of many of the presumed testing sites, standard laboratory equipment is impractical. A portable system is therefore a necessity.

Currently, two main types of sensing system are readily available for purchase.

The first is a strip test system. The great advantages of this system is that it is far cheaper than the alternative and highly portable – the strips are extremely light and can easily be carried in pockets if more than one site within a location is to be tested. It is also quick and easy to acquire. Searching the internet for ‘groundwater testing kit’ brings up dozens of websites from which these systems can be purchased instantly and in the possession of the customer within days.

The downsides to this system are that:

  1. the tests are disposable and suitable for a single use only meaning that many have to be used and the product continuously repurchased if long term testing is going to be considered.
  2. each strip will only test for one contaminant. Where a groundwater source is contaminated, it is unlikely that only a single potential contaminant will be of interest. For highly focussed research this may be acceptable, but in the case of broader research more than one kind of test strip will almost certainly be needed, and as with the problem of disposability, this will push up the costs.
  3. These tests are often not highly quantitative – quantitative to the extent of orders of magnitude but without providing precise information about contamination levels which makes them unsuitable for detailed examination of contamination levels at a source, especially if the study involves taking multiple readings over a period of time, when the change could be small.
A comparison of the ARSOlux kit (160 assays) (left) - a bioreporter kit - and the Arsonator kit (60 assays) (right) - an electronic water testing system. From Siegfried et al. Environ. Sci. Technol. 2012

The other major option is an electronic water testing system. These have the advantages of being far more quantitative, largely reusable and often considering more than one contaminant within the same system. As with the strip tests however, there are major disadvantages.

  1. acquiring these systems is not as easy. Often, instead of prices being offered, a quote must be obtained. A level of customisation is sometimes possible with the technology but it is invariably by far the more expensive of the two options.
  2. electronic systems tend to be bigger and heavier, with complete systems often coming in bulky cases that would be difficult to transport without a vehicle. These systems act more as a field lab, and as such portability is vastly reduced.

The major disadvantages to these systems are on one side a lack of quantitative results and reusability and on the other high, often prohibitive, costs.


How our system is different

Our system has been designed to be relatively low cost and the prices we have paid in producing the project, while not high, would be dramatically reduced by mass production. The system is light and portable but, between the output system and the implementation system, provides highly quantitative data. While the cuvettes and bacteria are single use, the implementation system is reusable.

We consider there to be a potential market for a reliable, accurate, precise, quantitative measuring system that without the price tag of current electronic systems. We also feel that our system could fill this niche.