Difference between revisions of "Team:Michigan/Experiments"

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   <div class = "container"> <h1 style="text-align:center; font-size: 75px;"><font face= "Poiret One">Public Engagement</font></h1>
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           <p style="text-align:center; font-size:20px;"><font face="verdana"> Synthetic biology has a long way to being completely accepted by the public, mainly due to lack of education about the subject. The Michigan Synthetic Biology Team held a few workshops in order to excite and incite younger generations to pursue synthetic biology!</font></p>
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           <p style="text-align:center; font-size:20px;"><font face="verdana"> Below we have outlined the path our project took, from the initial cloning through experimentation and submission.</font></p>
 
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        <img src="https://static.igem.org/mediawiki/2016/e/e0/T--Michigan--Detroit.jpg" width="500" height="340" style="float:left"><img src="https://static.igem.org/mediawiki/2016/6/68/T--Michigan--viruscards.jpg" width="500" height="340" style="float:right"></p></div>
 
 
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           <h1 style="text-align:center; font-size: 50px;"><font face= "Poiret One">Ann Arbor Health Hacks</h2>
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           <h1 style="text-align:center; font-size: 50px;"><font face= "Poiret One">Transferring LacZ into pET28 </h2>
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        <p style="text-align:center; font-size:20px;"><font face="verdana">We competed (and won a $500 award) in a health hackathon organized by Ann Arbor Health Hacks from June 14-16, 2016. Hackathons like these bring people with diverse backgrounds together to solve problems that they propose. We took our basic Aptapaper idea and through talking with the other hackathon participants worked to envision a version that could help diagnose heart disease.The hackathon was actually where we came up with the idea to use proximity dependent ligation instead of a toehold switch for our design. As part of the hackathon, we interacted with the other 200 or so participants from all different health related fields, teaching them about synthetic biology as they taught us about aspects of their particular field that were relevant to our project.</font></p>
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        <p style="text-align:center; font-size:20px;"><font face="verdana"> 1) Digested pET28 and new version of BBa_K564012 (with added cut sites)
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2) Ligated together
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3) Submitted for sequencing to confirm
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4) Transformed into new DH5a chemically competent cells
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         <h1 style="text-align:center; font-size: 50px;"> <font face= "Poiret One">Girls in Science and Engineering (GISE)</h2>
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         <h1 style="text-align:center; font-size: 50px;"> <font face= "Poiret One">Creation of delta-M15 mutant</h2>
 
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         <p style="text-align:center; font-size:20px;"><font face="verdana"> 1) Deletion of segment including alpha fragment of LacZ using NEB Q5 site directed mutagenesis kit
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<p style="text-align:center; font-size:20px;"><font face="verdana">In mid-June, the team helped out with the University of Michigan's GISE summer camp. We held two sessions where we showed middle-school girls how to extract DNA from strawberries and their own spit--an integral technique in synthetic biology! Aside from the activities, we explained the basics of genetics and synthetic biology. Despite the numerous spills and messes, the girls were intrigued with how the spidery liquid (full of their own DNA) contained in the microcentrifuge tube held all the instructions for their bodies! Hopefully, they'll be the next generation of Michigan Synthetic Biologists!</p>
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2) Submitted for sequencing to confirm successful deletion
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          <h1 style="text-align:center; font-size: 50px;"><font face= "Poiret One">Proximity dependent ligation assay </h2>
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        <p style="text-align:center; font-size:20px;"><font face="verdana"> 1) Probe segments and bridge segments mixed with T4 ligase in presence or absence of thrombin
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2) Gel electropheresis to detect ligation
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        <h1 style="text-align:center; font-size: 50px;"> <font face= "Poiret One">Cloned delta-M15 mutant into submission vector
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        <p style="text-align:center; font-size:20px;"><font face="verdana"> 1) Digested linearized pSB1C3 with our mutant
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2) Ligated together
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<p style="text-align:center; font-size:20px;"><font face="verdana"> 3) Transformed into new DH5a chemically competent cells
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5) Extracted DNA
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<p style="text-align:center; font-size:20px;"><font face="verdana"> 6) Dried, packaged, shipped </p>
 
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Revision as of 23:45, 19 October 2016


Home Project Modeling Team BioBrick Human Practices Safety Awards

Experimental Flow

Below we have outlined the path our project took, from the initial cloning through experimentation and submission.

Isolated LacZ from iGem part BBa_K564012

1) Rehydrated DNA from distribution kit

2) Transformed into DH5a chemically competent cells

3) Extracted plasmid

4) PCR mutagenesis to engineer desired cut sites around LacZ

Transferring LacZ into pET28

1) Digested pET28 and new version of BBa_K564012 (with added cut sites)

2) Ligated together

3) Submitted for sequencing to confirm

4) Transformed into new DH5a chemically competent cells

Creation of delta-M15 mutant


1) Deletion of segment including alpha fragment of LacZ using NEB Q5 site directed mutagenesis kit

2) Submitted for sequencing to confirm successful deletion

Proximity dependent ligation assay

1) Probe segments and bridge segments mixed with T4 ligase in presence or absence of thrombin

2) Gel electropheresis to detect ligation

Cloned delta-M15 mutant into submission vector


1) Digested linearized pSB1C3 with our mutant

2) Ligated together

3) Transformed into new DH5a chemically competent cells

5) Extracted DNA

6) Dried, packaged, shipped