Team:Auckland MOD/Notebook

<!DOCTYPE html> MOD - University of Auckland iGEM 2018 Team

Notebook

It's been a busy year! Take a look at the progress we've been making in our project: both in and outside of the lab.

Recruitment: We Want You!

As everyone comes back after the summer break, we begin recruiting new 2018 Team MOD members. The handover process from 2017 iGEM Team SECA means we're slowly building an iGEM alumni here at UoA. They've been really helpful mentoring us as we start our new project in terms of sharing research documentation, transferring financial accounts, and having a small team cheering for us from the sidelines.

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Identifying a National Problem

As a team we've all come together to discuss potential project ideas. Everyone has their own specific interests ranging from plant physiology to wine microbiology - it's great to see a diverse range of options. We've identified waterway pollution as a huge problem in New Zealand: each year we're seeing more incidences of toxic algal bloom formation in the summer and have begun searching potential ways to use synthetic biology to combat this issue.

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Refining our Biological Solution

After carrying out our own research, we've looked into the effects of dairy run-off entering the water system. Discussion with our PI and researchers within the university has helped narrow down the chosen metabolic pathway we might upregulate. We've chosen Arabidopsis thaliana as our model organism and plan on increasing the uptake and breakdown of excess urea in the soil, using the enzymes DUR3, UreG and GS1;2.

Lab induction time! We've all had to complete our safety training so we can jump into the PC1 facilities for our lab work.

Looking Beyond the Lab

If we're looking at a product for dairy farmers, it's important to start thinking about market validation. Rachel has begun speaking to social scientists in the university to get an idea of how to conduct a focus group, and how to engage with students as part of our community engagement efforts.

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Lorem ipsum dolor sit amet, consectetur adipiscing elit. Quisque iaculis nibh ut quam pellentesque eleifend. Quisque posuere dolor vitae quam tincidunt placerat. Suspendisse ex purus, ornare ut porta nec, interdum id diam. Morbi at mauris imperdiet, porttitor libero in, mattis magna. Pellentesque in mi tristique, congue ligula vitae, ultrices odio. Donec ac mauris nulla. Nam eu purus sed turpis congue placerat. Nunc ut lorem eu arcu auctor sollicitudin.

Thinking about our Business Model

Spent the entire week working towards our Return on Science presentation, really diving into the commercial aspects of our project and thinking about GMO policy in New Zealand which is exciting! We’ve also spoken to our secondary PI, Shaun Lott, about the future of iGEM in terms of next year’s team and getting recognition from the university as a contribution to the BSc degree.

Getting started with our BioBrick Characterisation

Week 22: Had a meeting with 2017 iGEM team lab members, Judith and Jess, to get a better idea of how we can work more efficiently. They suggested not to continue from their BioBrick project this year, but instead do a fluorescent or chromoprotein. We've chosen BBa_E0020 from 2004 - we’ll be finding out at what pH we observe the highest fluorescent intensity.

Week 23: We've met with our lab supervisor, Jasper, to go over our lab protocol and familiarise ourselves to the BioBrick standard system.

Week 24: Finally jumping into the lab to do our plating and preparatory steps for the transformation into competent E. coli strain, DH5α. We’re all starting to feel the pressure of upcoming exams which is getting in the way of lab progress. Right now, the priority is to grow our transformed E. coli and get them on ice so we can experiment on them later in July.

Week 25: Our first transformation of our BioBrick has been successful! We’re now waiting for our primers (from our IDT free DNA supply) to arrive in the mail so we can run colony PCR and confirm our insert is being expressed.

Designing our Urea Gene Constructs

We’re starting to design our GS1;2 construct to transform into Agrobacterium, with help of our PI and other researchers, Nijat Imin and Matthew Mayo-Smith. We plan on using the Gateway reaction (using the attB sites) to introduce our gene constructs into our chosen destination vectors. From their advice, we'll be using pDONR221 as our entry vector and pB2GW7 as our destination vector.

The UreG plasmid the plant lab ordered from another lab group arrived in the mail. We want to test our Gateway protocol as we wait for our IDT vectors to arrive, but first we need to find out how to add the attB sites to our plasmid.

Starting our Investor Presentations!

Presented our research to BioMatters! They were really receptive towards our idea in terms of it being relevant to nitrogen pollution in New Zealand. Such a cool experience connecting with people in the industry who are equally passionate about GMOs and are keen to support us!

Had our presentation with Return On Science! It’s cool to see our lab students present the entrepreneurial aspects of our project for the first time. We got some good feedback on areas of commercialization we should deeper look into - particularly around IP and our business model.

Working through the BioBrick lab

Week 27: Received our BioBrick primers from IDT and will begin the characterisation part of our project. It would be a good idea for future iGEM teams to provide primers for the BioBrick standard in the Distribution Kit. We’re also contacting past iGEM teams that have contributed to the same BioBrick or used similar protocols that we plan on using.

Week 28: We’ve run into some roadblocks in the BioBrick characterisation lab - spending the week troubleshooting our PCR protocol. We’ve found the polymerase we’ve been using hasn’t been that efficient, now using KAPA2G. Also looking at the BioBrick construct, our original Brick (BBa_E0020) doesn’t have a promoter so there’s no way to observe CFP expression unless we subclone it into another vector - (this seems like poor design in terms of iGEM’s standard system). For this reason we’ve switched to characterizing the RFP in the composite BBa_I3241 Brick.

Week 29: We’ve successfully transformed our composite RFP BioBrick into E. coli and run colony PCR to confirm. We’ll be spending the rest of the week planning our purification technique (subcloning into another vector to add a HisTag) and fluorescent characterisation under different salinities. We’ve found the transformed E. coli grow a bit slower than usual - I’ve sent out emails asking past teams that have worked with our BioBrick if they’ve encountered the same problems. Still waiting for them to reply!

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Lorem ipsum dolor sit amet, consectetur adipiscing elit. Quisque iaculis nibh ut quam pellentesque eleifend. Quisque posuere dolor vitae quam tincidunt placerat. Suspendisse ex purus, ornare ut porta nec, interdum id diam. Morbi at mauris imperdiet, porttitor libero in, mattis magna. Pellentesque in mi tristique, congue ligula vitae, ultrices odio. Donec ac mauris nulla. Nam eu purus sed turpis congue placerat. Nunc ut lorem eu arcu auctor sollicitudin.

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Lorem ipsum dolor sit amet, consectetur adipiscing elit. Quisque iaculis nibh ut quam pellentesque eleifend. Quisque posuere dolor vitae quam tincidunt placerat. Suspendisse ex purus, ornare ut porta nec, interdum id diam. Morbi at mauris imperdiet, porttitor libero in, mattis magna. Pellentesque in mi tristique, congue ligula vitae, ultrices odio. Donec ac mauris nulla. Nam eu purus sed turpis congue placerat. Nunc ut lorem eu arcu auctor sollicitudin.

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Lorem ipsum dolor sit amet, consectetur adipiscing elit. Quisque iaculis nibh ut quam pellentesque eleifend. Quisque posuere dolor vitae quam tincidunt placerat. Suspendisse ex purus, ornare ut porta nec, interdum id diam. Morbi at mauris imperdiet, porttitor libero in, mattis magna. Pellentesque in mi tristique, congue ligula vitae, ultrices odio. Donec ac mauris nulla. Nam eu purus sed turpis congue placerat. Nunc ut lorem eu arcu auctor sollicitudin.

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Lorem ipsum dolor sit amet, consectetur adipiscing elit. Quisque iaculis nibh ut quam pellentesque eleifend. Quisque posuere dolor vitae quam tincidunt placerat. Suspendisse ex purus, ornare ut porta nec, interdum id diam. Morbi at mauris imperdiet, porttitor libero in, mattis magna. Pellentesque in mi tristique, congue ligula vitae, ultrices odio. Donec ac mauris nulla. Nam eu purus sed turpis congue placerat. Nunc ut lorem eu arcu auctor sollicitudin.

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Lorem ipsum dolor sit amet, consectetur adipiscing elit. Quisque iaculis nibh ut quam pellentesque eleifend. Quisque posuere dolor vitae quam tincidunt placerat. Suspendisse ex purus, ornare ut porta nec, interdum id diam. Morbi at mauris imperdiet, porttitor libero in, mattis magna. Pellentesque in mi tristique, congue ligula vitae, ultrices odio. Donec ac mauris nulla. Nam eu purus sed turpis congue placerat. Nunc ut lorem eu arcu auctor sollicitudin.

Title

Lorem ipsum dolor sit amet, consectetur adipiscing elit. Quisque iaculis nibh ut quam pellentesque eleifend. Quisque posuere dolor vitae quam tincidunt placerat. Suspendisse ex purus, ornare ut porta nec, interdum id diam. Morbi at mauris imperdiet, porttitor libero in, mattis magna. Pellentesque in mi tristique, congue ligula vitae, ultrices odio. Donec ac mauris nulla. Nam eu purus sed turpis congue placerat. Nunc ut lorem eu arcu auctor sollicitudin.

Subtitle

Lorem ipsum dolor sit amet, consectetur adipiscing elit. Quisque iaculis nibh ut quam pellentesque eleifend. Quisque posuere dolor vitae quam tincidunt placerat. Suspendisse ex purus, ornare ut porta nec, interdum id diam. Morbi at mauris imperdiet, porttitor libero in, mattis magna. Pellentesque in mi tristique, congue ligula vitae, ultrices odio. Donec ac mauris nulla. Nam eu purus sed turpis congue placerat. Nunc ut lorem eu arcu auctor sollicitudin.

Title

Lorem ipsum dolor sit amet, consectetur adipiscing elit. Quisque iaculis nibh ut quam pellentesque eleifend. Quisque posuere dolor vitae quam tincidunt placerat. Suspendisse ex purus, ornare ut porta nec, interdum id diam. Morbi at mauris imperdiet, porttitor libero in, mattis magna. Pellentesque in mi tristique, congue ligula vitae, ultrices odio. Donec ac mauris nulla. Nam eu purus sed turpis congue placerat. Nunc ut lorem eu arcu auctor sollicitudin.

Subtitle

Lorem ipsum dolor sit amet, consectetur adipiscing elit. Quisque iaculis nibh ut quam pellentesque eleifend. Quisque posuere dolor vitae quam tincidunt placerat. Suspendisse ex purus, ornare ut porta nec, interdum id diam. Morbi at mauris imperdiet, porttitor libero in, mattis magna. Pellentesque in mi tristique, congue ligula vitae, ultrices odio. Donec ac mauris nulla. Nam eu purus sed turpis congue placerat. Nunc ut lorem eu arcu auctor sollicitudin.

"Competing in iGEM required more strength, endurance, and sacrifice than any other project I have undertaken. But nothing could have prepared me more for the post graduate study I am now pursuing than formulating and completing our project as an independent scienctist."

Judith Glasson, Alumni

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