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<h3>Designing our gene constructs</h3> | <h3>Designing our gene constructs</h3> | ||
− | <p>We’re starting to design our GS1;2 construct to transform into Agrobacterium, with help of our PI and other researchers in SBS. We plan on using the Gateway reaction ( | + | <p>We’re starting to design our GS1;2 construct to transform into Agrobacterium, with help of our PI and other researchers in SBS. We plan on using the Gateway reaction (using the <i>attB</i> sites) to introduce our gene constructs into our chosen destination vectors. From their advice, we'll be using <b>pDONR221</b> as our entry vector and <b>pB2GW7</b> as our destination vector. </p> |
− | <p> 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. </p> | + | <p> 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 <i>attB</i> sites to our plasmid. </p> |
<img src="https://static.igem.org/mediawiki/2018/f/f3/T--Auckland_MOD--lab.jpg"> | <img src="https://static.igem.org/mediawiki/2018/f/f3/T--Auckland_MOD--lab.jpg"> | ||
<h3>Starting our investor presentations!</h3> | <h3>Starting our investor presentations!</h3> | ||
<p> Had our first investor presentation with 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! </p> | <p> Had our first investor presentation with 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! </p> | ||
− | <p>Had our | + | <p>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. </p> |
<img src="https://2018.igem.org/File:T--Auckland_MOD--BiomattersPresentation.jpeg"> | <img src="https://2018.igem.org/File:T--Auckland_MOD--BiomattersPresentation.jpeg"> | ||
<h4>Working through the BioBrick lab</h4> | <h4>Working through the BioBrick lab</h4> | ||
<p> <b> Week 27: </b> Received our BioBrick primers from IDT and will begin the characterization 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. </p> | <p> <b> Week 27: </b> Received our BioBrick primers from IDT and will begin the characterization 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. </p> | ||
− | <p> <b> Week 28:</b> We’ve run into some roadblocks in the BioBrick characterization 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. </p> | + | <p> <b> Week 28:</b> We’ve run into some roadblocks in the BioBrick characterization 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 (<b>BBa_E0020</b>) 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 <b>BBa_I3241</b> Brick. </p> |
<p> <b> Week 29: </b> 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!</p> | <p> <b> Week 29: </b> 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!</p> | ||
</div> | </div> |
Revision as of 06:08, 24 July 2018
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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.
"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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