Team:RHIT/Notebook
From 2012.igem.org
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- | <img src="https://static.igem.org/mediawiki/igem.org/8/87/Banner_only2.png" width="100%"/> | + | <a href="https://2012.igem.org/Team:RHIT"><img src="https://static.igem.org/mediawiki/igem.org/8/87/Banner_only2.png" width="100%"/></a> |
<div id="logged_out"> | <div id="logged_out"> | ||
<a href="https://2012.igem.org/Main_Page">iGEM Home</a> | <a href="https://2012.igem.org/Main_Page">iGEM Home</a> | ||
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<div class="rhit-blueSheet" id="rhit-blueSheet"> | <div class="rhit-blueSheet" id="rhit-blueSheet"> | ||
<h4>Week 1</h4> | <h4>Week 1</h4> | ||
- | <ul id="dailyLog | + | <ul id="dailyLog"> |
<li>6/4/2012: First round of project planning, lab clean-up, general outline for summer.</li><br /> | <li>6/4/2012: First round of project planning, lab clean-up, general outline for summer.</li><br /> | ||
<li>6/5/2012: Second round of project planning, lab clean-up, research into Johns Hopkins 2008 summer project.</li><br /> | <li>6/5/2012: Second round of project planning, lab clean-up, research into Johns Hopkins 2008 summer project.</li><br /> | ||
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</ul> | </ul> | ||
<h4>Week 2</h4> | <h4>Week 2</h4> | ||
- | <ul id="dailyLog | + | <ul id="dailyLog"> |
<li>6/11/2012: Identified a positive feedback loop that's been done in yeast before and a possible selection scheme, started plasmid mapping, set up server for wiki, began wiki design and looked at color schemes, block-diagrammed positive feedback loop, ate a whole tray of brownies.</li><br /> | <li>6/11/2012: Identified a positive feedback loop that's been done in yeast before and a possible selection scheme, started plasmid mapping, set up server for wiki, began wiki design and looked at color schemes, block-diagrammed positive feedback loop, ate a whole tray of brownies.</li><br /> | ||
<li>6/12/2012: Found gene sequences, continued work on graphic for top of Wiki, looked for selection schemes, shot down selection schemes.</li><br /> | <li>6/12/2012: Found gene sequences, continued work on graphic for top of Wiki, looked for selection schemes, shot down selection schemes.</li><br /> | ||
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</ul> | </ul> | ||
<h4>Week 3</h4> | <h4>Week 3</h4> | ||
- | <ul id="dailyLog | + | <ul id="dailyLog"> |
<li>6/18/2012: Optimized DNA sequences, found all of the filler DNA.</li><br /> | <li>6/18/2012: Optimized DNA sequences, found all of the filler DNA.</li><br /> | ||
<li>6/19/2012: Talked with Dr. Almaas, started web design, took pictures with storm troopers, team movie night.</li><br /> | <li>6/19/2012: Talked with Dr. Almaas, started web design, took pictures with storm troopers, team movie night.</li><br /> | ||
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</ul> | </ul> | ||
<h4>Week 4</h4> | <h4>Week 4</h4> | ||
- | <ul id="dailyLog | + | <ul id="dailyLog"> |
<li>6/25/2012: Group discussion, work on school website, organization of collected information.</li><br /> | <li>6/25/2012: Group discussion, work on school website, organization of collected information.</li><br /> | ||
<li>6/26/2012: Obtained shirts, found and organized all sequences, started to verify sequences, school page is 85% done.</li><br /> | <li>6/26/2012: Obtained shirts, found and organized all sequences, started to verify sequences, school page is 85% done.</li><br /> | ||
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<h4>Team Break Week</h4> | <h4>Team Break Week</h4> | ||
<h4>Week 5</h4> | <h4>Week 5</h4> | ||
- | <ul id="dailyLog | + | <ul id="dailyLog"> |
<li>7/9/2012: Got back from break. Learned DNA had not shipped yet. :( Worked on wiki.</li><br /> | <li>7/9/2012: Got back from break. Learned DNA had not shipped yet. :( Worked on wiki.</li><br /> | ||
<li>7/10/2012: Worked on wiki.</li><br /> | <li>7/10/2012: Worked on wiki.</li><br /> | ||
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</ul> | </ul> | ||
<h4>Week 6</h4> | <h4>Week 6</h4> | ||
- | <ul id="dailyLog | + | <ul id="dailyLog"> |
<li>7/16/2012: Decided to use GeneArt for sequences instead of original DNA company.</li><br /> | <li>7/16/2012: Decided to use GeneArt for sequences instead of original DNA company.</li><br /> | ||
<li>7/18/2012: Brainstormed ideas for side projects while waiting for DNA shipment.</li><br /> | <li>7/18/2012: Brainstormed ideas for side projects while waiting for DNA shipment.</li><br /> | ||
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</ul> | </ul> | ||
<h4>Week 7</h4> | <h4>Week 7</h4> | ||
- | <ul id="dailyLog | + | <ul id="dailyLog"> |
<li>7/23/2012: Devon and Kristen began planning the THCM exhibit.</li><br /> | <li>7/23/2012: Devon and Kristen began planning the THCM exhibit.</li><br /> | ||
<li>7/24/2012: Continued work in Teams. Began intial research into pathway for modeling.</li><br /> | <li>7/24/2012: Continued work in Teams. Began intial research into pathway for modeling.</li><br /> | ||
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</ul> | </ul> | ||
<h4>Week 8</h4> | <h4>Week 8</h4> | ||
- | <ul id="dailyLog | + | <ul id="dailyLog"> |
<li>7/30/2012: Ironed out wiki formatting, began adding content, continued Maya animation.</li><br /> | <li>7/30/2012: Ironed out wiki formatting, began adding content, continued Maya animation.</li><br /> | ||
<li>7/31/2012: Lab work, continued adding content to wiki.</li><br /> | <li>7/31/2012: Lab work, continued adding content to wiki.</li><br /> | ||
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</ul> | </ul> | ||
<h4>Week 9</h4> | <h4>Week 9</h4> | ||
- | <ul id="dailyLog | + | <ul id="dailyLog"> |
<li>8/6/2012: Canoe trip with IRC members, lab work.</li><br /> | <li>8/6/2012: Canoe trip with IRC members, lab work.</li><br /> | ||
<li>8/7/2012: Lab work, focus on modeling information for NTNU, continued work in Teams.</li><br /> | <li>8/7/2012: Lab work, focus on modeling information for NTNU, continued work in Teams.</li><br /> | ||
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</ul> | </ul> | ||
<h4>Week 10</h4> | <h4>Week 10</h4> | ||
- | <ul id="dailyLog | + | <ul id="dailyLog"> |
<li>8/13/2012: Lab work, continued work in Teams.</li><br /> | <li>8/13/2012: Lab work, continued work in Teams.</li><br /> | ||
<li>8/14/2012: Lab work, continued work in Teams.</li><br /> | <li>8/14/2012: Lab work, continued work in Teams.</li><br /> | ||
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<h3>September 30, 2012</h3> | <h3>September 30, 2012</h3> | ||
Performed Mini-preps of transformed E.Coli, Began yeast transformation, plated additional yeast cultures.<br /> | Performed Mini-preps of transformed E.Coli, Began yeast transformation, plated additional yeast cultures.<br /> | ||
+ | Members: Adam | ||
+ | <h3>October 1, 2012</h3> | ||
+ | Plated Yeast Transformants<br /> | ||
+ | Members: Adam | ||
+ | <h3>October 3, 2012</h3> | ||
+ | Transformed yeast found growing on plates<br /> | ||
Members: Adam | Members: Adam | ||
</div> | </div> | ||
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<h2>Strains Used</h2> | <h2>Strains Used</h2> | ||
<h3><i>E. Coli</i></h3> | <h3><i>E. Coli</i></h3> | ||
- | <p>NEB5-alpha (NEB #C2988J or #C2987I): fhuA2 | + | <p>NEB5-alpha (NEB #C2988J or #C2987I): <i>fhuA2 ?(argF-lacZ)U169 phoA glnV44 F80? (lacZ)M15 gyrA96 recA1 relA1 endA1 thi-1 hsdR17</i></p><br /> |
<h3><i>S. Cerevisiae</i></h3> | <h3><i>S. Cerevisiae</i></h3> | ||
- | <p>BY4741 (ATCC #201388): <i>MATa his3- | + | <p>BY4741 (ATCC #201388): <i>MATa his3-?0 leu2-?0 met15-?0 ura3-?0</i></p><br /> |
- | <p>BY4742 (ATCC #201389): <i> | + | <p>BY4742 (ATCC #201389): <i>MATalpha his3-?0 leu2-?0 lys2-?0 ura3-?0</i></p><br /><br /> |
+ | <h2>Lab Protocols</h2> | ||
+ | <p>Transformations were carried out on competent E.Coli using the procedure available on the <a href="http://www.neb.com/nebecomm/products/protocol119.asp">NEB website</a>.</p><br /><br /> | ||
+ | |||
+ | <p>DNA extraction from cells was performed using a Qiagen QIAprep Spin Miniprep Kit (Cat. no. 27104). Included procedure was followed.</p><br /><br /> | ||
+ | |||
+ | <p>Digestions were performed using EcoRI, XbaI, SpeI and PstI using recommended NEB procedure.</p><br /><br /> | ||
+ | |||
+ | <p>Gel extraction of digested DNA was performed using a Qiagen QIAEX II Gel Extraction Kit (Cat. no. 20021). Included procedure was followed.</p><br /><br /> | ||
+ | |||
+ | <p>Ligations were performed following procedure available on the <a href="http://www.neb.com/nebecomm/products/protocol658.asp">NEB website</a>. Total mass of DNA for ligations was 100ng with a 1:3 to 1:4 ratio of vector to insert. </p><br /><br /> | ||
+ | |||
</div> | </div> | ||
<div class="rhit-fourthSheet" id="rhit-fourthSheet"> | <div class="rhit-fourthSheet" id="rhit-fourthSheet"> | ||
- | The official RHIT Safety page can be found <a href="https://2012.igem.org/Team:RHIT/Safety">here | + | The official RHIT Safety page can be found <a href="https://2012.igem.org/Team:RHIT/Safety">here</a>. |
<h2>Researcher, Public, and Environmental Safety</h2> | <h2>Researcher, Public, and Environmental Safety</h2> | ||
<p>The Rose-Hulman iGEM team has considered safety a top priority since the early stages of project development. | <p>The Rose-Hulman iGEM team has considered safety a top priority since the early stages of project development. | ||
As a result, projects that could pose a significant risk were dismissed. | As a result, projects that could pose a significant risk were dismissed. | ||
Our laboratory space is considered a Basic Biosafety Level 1 laboratory but it does have a few characteristics of higher categories. | Our laboratory space is considered a Basic Biosafety Level 1 laboratory but it does have a few characteristics of higher categories. | ||
- | It has controlled access, biohazard signs and waste disposal bins, and an autoclave. | + | It has controlled access, biohazard signs and waste disposal bins, and an autoclave. Other safety features are demonstrated below.<br /></p> |
- | All participating students received standard safety and good laboratory practice training as part of their academic laboratory course-work. | + | <div align="center"><img src="https://static.igem.org/mediawiki/igem.org/2/2a/Safety_pics.png" width="60%"></div><br /> |
+ | <p>All participating students received standard safety and good laboratory practice training as part of their academic laboratory course-work. | ||
The Instructor provided more specific safety instruction, as necessary. | The Instructor provided more specific safety instruction, as necessary. | ||
Proper attire was worn at all times in the laboratory. Gloves and glasses were donned as warranted. | Proper attire was worn at all times in the laboratory. Gloves and glasses were donned as warranted. | ||
- | The only organisms used in our project are common laboratory strains of < | + | The only organisms used in our project are common laboratory strains of <i>Saccharomyces cerevisiae</i> (BY4741 and BY4742) and <i>Escherichia coli</i> (NEB5alpa), |
which are both considered Risk Group 1 microorganisms according to the Laboratory Biosafety Manual published by the World Health Organization. | which are both considered Risk Group 1 microorganisms according to the Laboratory Biosafety Manual published by the World Health Organization. | ||
Group 1 organisms are defined as “no or low individual and community risk, a microorganism that is unlikely to cause human or animal disease.” | Group 1 organisms are defined as “no or low individual and community risk, a microorganism that is unlikely to cause human or animal disease.” |
Latest revision as of 03:57, 4 October 2012