Team:RHIT/Notebook

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var split = location.search.replace('?', '').split('=');

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<a href="https://2012.igem.org/wiki/index.php?title=Special:UserLogin&returnto=2012.igem.org/Team:RHIT">Login</a>

</div>

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-

+

-

<a id='account-link' class='RHIT-external' href='https://igem.org/User_Information'>My Account</a> 

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-

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<a id='account-link' class='RHIT-external' href='https://igem.org/User_Information'>My Account</a> 

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</div>

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</div>

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<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 />

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</ul>

-

+

<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 />

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</ul>

-

+

<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 />

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</ul>

-

+

<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 />

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<h4>Team Break Week</h4>

-

+

<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 />

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</ul>

-

+

<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 />

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</ul>

-

+

<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 />

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</ul>

-

+

<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>8/1/2012: Lab work, added more content to wiki, got update on additional computer for Maya animation, made significant progress on math model, Rose-Hulman team page went live.</li><br />

<li>8/2/2012: Lab work, continued work in Teams.</li><br />

-

<li>8/3/2012: Lab work, continued work in Teams.</li~~><br /~~>

+

<li>8/3/2012: Lab work, continued work in Teams.</li>

</ul>

-

+

<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>8/8/2012: Second Skype meeting with NTNU~~, continued work in teams~~, conclusion and review of Maya animation, ~~lab~~ work.</li><br />

+

<li>8/8/2012: Lab work, Second Skype meeting with NTNU, conclusion and review of Maya animation.</li><br />

+

<li>8/9/2012: Lab work, continued work in Teams.</li><br />

+

<li>8/10/2012: Lab work, continued work in Teams.</li>

+

</ul>

+

+

+

<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>8/15/2012: Lab work, continued work in Teams.</li><br />

+

<li>8/16/2012: Lab work, plans made for continuing into the school year.</li><br />

+

<li>8/17/2012: Cleaned workspace, finalized plans for continuation.</li><br />

</ul>

</div></a>

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Transformed E. coli with our red construct<br />

Members: Bobby, Adam, Devon, Kristen, Ben, Dr. A

+

<h3>August 8, 2012</h3>

+

Inoculated liquid media with transformed E.Coli<br />

+

Members: Bobby, Adam

+

<h3>August 9, 2012</h3>

+

Ran Mini-Preps of pRS413, pRS416, and Red construct bearing E. Coli, stored extracted DNA<br />

+

Members: Bobby, Adam

+

<h3>August 10, 2012</h3>

+

Ran quick gel of extracted DNA<br />

+

Members: Bobby, Adam

+

<h3>August 11, 2012</h3>

+

Digested extracted DNA, isolated pRS413 & pRS416 fragments from gel<br />

+

Members: Bobby, Adam

+

<h3>August 12, 2012</h3>

+

Isolated red construct fragments on gel, poured chloramphenicol plates<br />

+

Members: Bobby, Adam

+

<h3>August 13, 2012</h3>

+

Digested extracted DNA, isolated pRS413, pRS416, and Red Construct from gel, Transformed E. Coli with NTNU construct<br />

+

Members: Bobby, Adam, Dr. A

+

<h3>August 14, 2012</h3>

+

Ligated pRS413 and red construct, transformed E. Coli with ligated plasmid, inoculated liquid media with NTNU E. Coli<br />

+

Members: Bobby, Adam

+

<h3>August 15, 2012</h3>

+

Mini-Preps of NTNU construct, confirmed presence on gel, Digested pRS413, pRS416, and Red Construct, poured ampicillin plates<br />

+

Members: Bobby, Adam

+

<h3>August 16, 2012</h3>

+

Isolated pRS413, pRS416, and Red Construct from gel, began storing strains, DNA, and equipment<br />

+

Members: Bobby, Adam

+

<h3>August 17, 2012</h3>

+

Ran digestion of pRS413, pRS416, and Red Construct, stored remaining strains, DNA, and equipment<br />

+

Members: Bobby, Adam, Dr. A

+

<h3>August 28, 2012</h3>

+

Transformed E.Coli with BBa_E0422 reporter<br />

+

Members: Bobby, Adam, Dr. A

+

<h3>August 29, 2012</h3>

+

Digested and isolated Red Construct and pRS413 from gel, Inoculated liquid media with BBa_E0422 reporter<br />

+

Members: Adam

+

<h3>August 30, 2012</h3>

+

Digested pRS413 and Red Construct, Isolated from gel and Ligated, Transformed E. Coli.<br />

+

Members: Adam

+

<h3>August 31, 2012</h3>

+

Digested pRS413 and Red Construct<br />

+

Members: Bobby, Adam, Dr. A

+

<h3>September 1, 2012</h3>

+

Started cultures of pRS413 and the Red construct form frozen cultures<br />

+

Members: Adam

+

<h3>September 2, 2012</h3>

+

Inoculated liquid media with colonies from plates of pRS413 and Red Construct strains<br />

+

Members: Adam

+

<h3>September 3, 2012</h3>

+

Ran mini-preps of liquid colonies<br />

+

Members: Adam

+

<h3>September 4, 2012</h3>

+

Ran diagnostic gel to check amount of DNA extracted<br />

+

Members: Adam

+

<h3>September 5, 2012</h3>

+

Purified and concentrated DNA samples<br />

+

Members: Adam

+

<h3>September 6, 2012</h3>

+

Ran diagnostic gel to check for purification success<br />

+

Members: Adam

+

<h3>September 7, 2012</h3>

+

Ran BioBrick Assembly protocol on NTNU lld promoter and BBa_E0422, Transformed E.Coli with ligation mixture<br />

+

Members: Adam, Dr. A

+

<h3>September 8, 2012</h3>

+

Inoculated liquid cultures with transformed E.Coli<br />

+

Members: Adam

+

<h3>September 10, 2012</h3>

+

Digested and isolated Red Construct and pRS413 from gel, Ran mini-preps of liquid cultures<br />

+

Members: Adam

+

<h3>September 11, 2012</h3>

+

Ran gel of lld/0422 hybrid, Gel showed presence of desired product <br />

+

Members: Adam

+

<h3>September 12, 2012</h3>

+

Inoculated liquid cultures and plates with transformed E.Coli<br />

+

Members: Adam

+

<h3>September 13, 2012</h3>

+

Performed ligation on Red Construct, Transformed E.Coli with blue construct<br />

+

Members: Adam

+

<h3>September 14, 2012</h3>

+

Inoculated liquid cultures with blue construct transformants<br />

+

Members: Adam

+

<h3>September 15, 2012</h3>

+

Ran mini-preps on colonies with blue constructs<br />

+

Members: Adam and Bobby

+

<h3>September 16, 2012</h3>

+

Digested Blue construct and pRS413, isolated desired fragments from gel<br />

+

Members: Adam

+

<h3>September 17, 2012</h3>

+

Ligated Blue construct and pRS413 fragments, transformed E.Coli<br />

+

Members: Adam

+

<h3>September 18, 2012</h3>

+

Inoculated Liquid Media with Blue construct/pRS413 fusion bearing E.Coli<br />

+

Members: Adam

+

<h3>September 19, 2012</h3>

+

NTNU construct testing via fluorescence, Mini-Preps of Blue construct/pRS413 fusion bearing E.Coli<br />

+

Members: Bobby, Alex, Adam

+

<h3>September 20, 2012</h3>

+

Diagnostic gels of Blue Construct/pRS413 mini-preps<br />

+

Members: Adam

+

<h3>September 22, 2012</h3>

+

Inoculated liquid media with Blue Construct/pRS413 E. Coli, prepared stock solutions<br />

+

Members: Adam, Dr. A

+

<h3>September 23, 2012</h3>

+

Mini-Preps of E.Coli, Ran gel of prep, made liquid media<br />

+

Members: Adam

+

<h3>September 24, 2012</h3>

+

Ran digestion of mini-prep DNA, plated Yeast<br />

+

Members: Adam

+

<h3>September 25, 2012</h3>

+

Gel of digestion result, isolated fragments from gel<br />

+

Members: Adam

+

<h3>September 26, 2012</h3>

+

Ligated Blue Fragment/pRS413, transformed E.Coli<br />

+

Members: Adam

+

<h3>September 27, 2012</h3>

+

Inoculated LB-AMP broth with transformed E.Coli, plated Yeast<br />

+

Members: Adam

+

<h3>September 28, 2012</h3>

+

Inoculated LB-CARB broth with transformed E.Coli<br />

+

Members: Adam

+

<h3>September 29, 2012</h3>

+

Inoculated LB-CARB broth with transformed E.Coli<br />

+

Members: Adam

+

<h3>September 30, 2012</h3>

+

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

</div>

-

~~Protocol text~~

+

<h2>Strains Used</h2>

+

+

<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>

+

+

<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>

-

Safety ~~text~~

+

The official RHIT Safety page can be found <a href="https://2012.igem.org/Team:RHIT/Safety">here</a>.

-

</div>

+

<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.

+

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.

+

It has controlled access, biohazard signs and waste disposal bins, and an autoclave. Other safety features are demonstrated below.<br /></p>

+

+

<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.

+

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 <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.

+

Group 1 organisms are defined as “no or low individual and community risk, a microorganism that is unlikely to cause human or animal disease.”

+

Aseptic technique is used whenever working with these organisms and any contaminated wastes are sterilized by autoclaving or destroyed by commercial pyrolysis.

+

All laboratory chemicals are stored, handled and used as recommended by the manufacturer,

+

and they are disposed of in accordance with national, state, and local regulations and recommendations.

+

The laboratory space and contents are not accessible by unauthorized personnel.

+

All microbial strains, including bacteria rendered antibiotic resistant by transformation, harbor nutritional auxotrophies or other mutations

+

that mitigate the risk of their growing outside of the laboratory or causing disease in healthy humans or animals.

+

Furthermore, none of our recombinant constructs produce any known contagion or toxin.<p>

+

<h2>BioBrick Safety</h2>

+

<p>None of the BioBrick parts utilized or constructed are known to pose any safety issues.

+

Furthermore, they are well contained by the microbes that harbor them. The risk of unintended transfer to any other organism is minimal.</p>

+

<h2>Biosafety Group</h2>

+

<p>Rose-Hulman does not have a biosafety group, committee or review board other than an Animal Care and Use Committee,

+

which oversees animal research. Safety training and laboratory waste disposal are facilitated by an Environmental Health and Safety Officer, who also serves as a resource for faculty and students.</p>

+

<h2>Future iGEM Safety</h2>

+

<p>The possibility of designing mutually dependent strain/vector systems for routine manipulation and storage of BioBrick parts should be explored.</p>

+

</div>

</body>

-

~~</html>~~

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                                          $('#logged_in').show();
                                  }
+							$('#rhit-sheet2').hide();
+var split = location.search.replace('?', '').split('=');
+window[split[1]]();
 			});
-						function page1() {
+			function page1() {
 				$('#rhit-blueSheet').show();
 				$('#rhit-redSheet').hide();
@@ Line 44: / Line 47: @@
 		<div class="rhit-content">
 			<div class="rhit-header">
-				<img src="/wiki/images/f/f2/Rhit-banner.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">
+					<a href="https://2012.igem.org/Main_Page">iGEM Home</a>
 					<a href="https://2012.igem.org/wiki/index.php?title=Special:UserLogin&returnto=2012.igem.org/Team:RHIT">Login</a>
 				</div>
-                                <div id="logged_in">
+				<div id="logged_in">
-                                        <a id='user-link' class='RHIT-external' href='#'></a>&nbsp;
+					<a href="https://2012.igem.org/Main_Page">iGEM Home</a>
-                                        <a id='account-link' class='RHIT-external' href='https://igem.org/User_Information'>My Account</a>&nbsp;
+					<a id='user-link' class='RHIT-external' href='#'></a>&nbsp;
-                                        <a id='logout-link' class='RHIT-external' href='https://2012.igem.org/wiki/index.php?title=Special:UserLogout&returnto=2012.igem.org/Team:RHIT'>Log Out</a>
+					<a id='account-link' class='RHIT-external' href='https://igem.org/User_Information'>My Account</a>&nbsp;
-                                </div>
+					<a id='logout-link' class='RHIT-external' href='https://2012.igem.org/wiki/index.php?title=Special:UserLogout&returnto=2012.igem.org/Team:RHIT'>Log Out</a>
+				</div>
 			</div>
 			<div class="rhit-navLeft">
@@ Line 100: / Line 105: @@
 				<div class="rhit-blueSheet" id="rhit-blueSheet">
 					<h4>Week 1</h4>
-					<ul id="dailyLog" style="list-style-type:none">
+					<ul id="dailyLog">
 					<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 />
@@ Line 108: / Line 113: @@
 					</ul>
 					<h4>Week 2</h4>
-					<ul id="dailyLog" style="list-style-type:none">
+					<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>6/12/2012: Found gene sequences, continued work on graphic for top of Wiki, looked for selection schemes, shot down selection schemes.</li><br />
@@ Line 116: / Line 121: @@
 					</ul>
 					<h4>Week 3</h4>
-					<ul id="dailyLog" style="list-style-type:none">
+					<ul id="dailyLog">
 					<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 />
@@ Line 124: / Line 129: @@
 					</ul>
 					<h4>Week 4</h4>
-					<ul id="dailyLog" style="list-style-type:none">
+					<ul id="dailyLog">
 					<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 />
@@ Line 133: / Line 138: @@
 					<h4>Team Break Week</h4>
 					<h4>Week 5</h4>
-					<ul id="dailyLog" style="list-style-type:none">
+					<ul id="dailyLog">
 					<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 />
@@ Line 141: / Line 146: @@
 					</ul>
 					<h4>Week 6</h4>
-					<ul id="dailyLog" style="list-style-type:none">
+					<ul id="dailyLog">
 					<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 />
@@ Line 149: / Line 154: @@
 					</ul>
 					<h4>Week 7</h4>
-					<ul id="dailyLog" style="list-style-type:none">
+					<ul id="dailyLog">
 					<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 />
@@ Line 157: / Line 162: @@
 					</ul>
 					<h4>Week 8</h4>
-					<ul id="dailyLog" style="list-style-type:none">
+					<ul id="dailyLog">
 					<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>8/1/2012: Lab work, added more content to wiki, got update on additional computer for Maya animation, made significant progress on math model, Rose-Hulman team page went live.</li><br />
 					<li>8/2/2012: Lab work, continued work in Teams.</li><br />
-					<li>8/3/2012: Lab work, continued work in Teams.</li><br />
+					<li>8/3/2012: Lab work, continued work in Teams.</li>
 					</ul>
 					<h4>Week 9</h4>
-					<ul id="dailyLog" style="list-style-type:none">
+					<ul id="dailyLog">
 					<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>8/8/2012: Second Skype meeting with NTNU, continued work in teams, conclusion and review of Maya animation, lab work.</li><br />
+					<li>8/8/2012: Lab work, Second Skype meeting with NTNU, conclusion and review of Maya animation.</li><br />
+					<li>8/9/2012: Lab work, continued work in Teams.</li><br />
+					<li>8/10/2012: Lab work, continued work in Teams.</li>
+					</ul>
+					<h4>Week 10</h4>
+					<ul id="dailyLog">
+					<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>8/15/2012: Lab work, continued work in Teams.</li><br />
+					<li>8/16/2012: Lab work, plans made for continuing into the school year.</li><br />
+					<li>8/17/2012: Cleaned workspace, finalized plans for continuation.</li><br />
 					</ul>
 				</div></a>
@@ Line 190: / Line 205: @@
 					Transformed E. coli with our red construct<br />
 					Members: Bobby, Adam, Devon, Kristen, Ben, Dr. A
+					<h3>August 8, 2012</h3>
+					Inoculated liquid media with transformed E.Coli<br />
+					Members: Bobby, Adam
+					<h3>August 9, 2012</h3>
+					Ran Mini-Preps of pRS413, pRS416, and Red construct bearing E. Coli, stored extracted DNA<br />
+					Members: Bobby, Adam
+					<h3>August 10, 2012</h3>
+					Ran quick gel of extracted DNA<br />
+					Members: Bobby, Adam
+					<h3>August 11, 2012</h3>
+					Digested extracted DNA, isolated pRS413 & pRS416 fragments from gel<br />
+					Members: Bobby, Adam
+					<h3>August 12, 2012</h3>
+					Isolated red construct fragments on gel, poured chloramphenicol plates<br />
+					Members: Bobby, Adam
+					<h3>August 13, 2012</h3>
+					Digested extracted DNA, isolated pRS413, pRS416, and Red Construct from gel, Transformed E. Coli with NTNU construct<br />
+					Members: Bobby, Adam, Dr. A
+					<h3>August 14, 2012</h3>
+					Ligated pRS413 and red construct, transformed E. Coli with ligated plasmid, inoculated liquid media with NTNU E. Coli<br />
+					Members: Bobby, Adam
+					<h3>August 15, 2012</h3>
+					Mini-Preps of NTNU construct, confirmed presence on gel, Digested pRS413, pRS416, and Red Construct, poured ampicillin plates<br />
+					Members: Bobby, Adam
+					<h3>August 16, 2012</h3>
+					Isolated pRS413, pRS416, and Red Construct from gel, began storing strains, DNA, and equipment<br />
+					Members: Bobby, Adam
+					<h3>August 17, 2012</h3>
+					Ran digestion of pRS413, pRS416, and Red Construct, stored remaining strains, DNA, and equipment<br />
+					Members: Bobby, Adam, Dr. A
+					<h3>August 28, 2012</h3>
+					Transformed E.Coli with BBa_E0422 reporter<br />
+					Members: Bobby, Adam, Dr. A
+					<h3>August 29, 2012</h3>
+					Digested and isolated Red Construct and pRS413 from gel, Inoculated liquid media with BBa_E0422 reporter<br />
+					Members: Adam
+					<h3>August 30, 2012</h3>
+					Digested pRS413 and Red Construct, Isolated from gel and Ligated, Transformed E. Coli.<br />
+					Members: Adam
+					<h3>August 31, 2012</h3>
+					Digested pRS413 and Red Construct<br />
+					Members: Bobby, Adam, Dr. A
+					<h3>September 1, 2012</h3>
+					Started cultures of pRS413 and the Red construct form frozen cultures<br />
+					Members: Adam
+					<h3>September 2, 2012</h3>
+					Inoculated liquid media with colonies from plates of pRS413 and Red Construct strains<br />
+					Members: Adam
+					<h3>September 3, 2012</h3>
+					Ran mini-preps of liquid colonies<br />
+					Members: Adam
+					<h3>September 4, 2012</h3>
+					Ran diagnostic gel to check amount of DNA extracted<br />
+					Members: Adam
+					<h3>September 5, 2012</h3>
+					Purified and concentrated DNA samples<br />
+					Members: Adam
+					<h3>September 6, 2012</h3>
+					Ran diagnostic gel to check for purification success<br />
+					Members: Adam
+					<h3>September 7, 2012</h3>
+					Ran BioBrick Assembly protocol on NTNU lld promoter and BBa_E0422, Transformed E.Coli with ligation mixture<br />
+					Members: Adam, Dr. A
+					<h3>September 8, 2012</h3>
+					Inoculated liquid cultures with transformed E.Coli<br />
+					Members: Adam
+					<h3>September 10, 2012</h3>
+					Digested and isolated Red Construct and pRS413 from gel, Ran mini-preps of liquid cultures<br />
+					Members: Adam
+					<h3>September 11, 2012</h3>
+					Ran gel of lld/0422 hybrid, Gel showed presence of desired product <br />
+					Members: Adam
+					<h3>September 12, 2012</h3>
+					Inoculated liquid cultures and plates with transformed E.Coli<br />
+					Members: Adam
+					<h3>September 13, 2012</h3>
+					Performed ligation on Red Construct, Transformed E.Coli with blue construct<br />
+					Members: Adam
+					<h3>September 14, 2012</h3>
+					Inoculated liquid cultures with blue construct transformants<br />
+					Members: Adam
+					<h3>September 15, 2012</h3>
+					Ran mini-preps on colonies with blue constructs<br />
+					Members: Adam and Bobby
+					<h3>September 16, 2012</h3>
+					Digested Blue construct and pRS413, isolated desired fragments from gel<br />
+					Members: Adam
+					<h3>September 17, 2012</h3>
+					Ligated Blue construct and pRS413 fragments, transformed E.Coli<br />
+					Members: Adam
+					<h3>September 18, 2012</h3>
+					Inoculated Liquid Media with Blue construct/pRS413 fusion bearing E.Coli<br />
+					Members: Adam
+					<h3>September 19, 2012</h3>
+					NTNU construct testing via fluorescence, Mini-Preps of Blue construct/pRS413 fusion bearing E.Coli<br />
+					Members: Bobby, Alex, Adam
+					<h3>September 20, 2012</h3>
+					Diagnostic gels of Blue Construct/pRS413 mini-preps<br />
+					Members: Adam
+					<h3>September 22, 2012</h3>
+					Inoculated liquid media with Blue Construct/pRS413 E. Coli, prepared stock solutions<br />
+					Members: Adam, Dr. A
+					<h3>September 23, 2012</h3>
+					Mini-Preps of E.Coli, Ran gel of prep, made liquid media<br />
+					Members: Adam
+					<h3>September 24, 2012</h3>
+					Ran digestion of mini-prep DNA, plated Yeast<br />
+					Members: Adam
+					<h3>September 25, 2012</h3>
+					Gel of digestion result, isolated fragments from gel<br />
+					Members: Adam
+					<h3>September 26, 2012</h3>
+					Ligated Blue Fragment/pRS413, transformed E.Coli<br />
+					Members: Adam
+					<h3>September 27, 2012</h3>
+					Inoculated LB-AMP broth with transformed E.Coli, plated Yeast<br />
+					Members: Adam
+					<h3>September 28, 2012</h3>
+					Inoculated LB-CARB broth with transformed E.Coli<br />
+					Members: Adam
+					<h3>September 29, 2012</h3>
+					Inoculated LB-CARB broth with transformed E.Coli<br />
+					Members: Adam
+					<h3>September 30, 2012</h3>
+					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
 				</div>
 				<div class="rhit-grnSheet" id="rhit-grnSheet">
-					Protocol text
+					<h2>Strains Used</h2>
+					<h3><i>E. Coli</i></h3>
+					<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>
+					<p>BY4741 (ATCC #201388): <i>MATa his3-?0 leu2-?0 met15-?0 ura3-?0</i></p><br />
+					<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 class="rhit-fourthSheet" id="rhit-fourthSheet">
-					Safety text
+					The official RHIT Safety page can be found <a href="https://2012.igem.org/Team:RHIT/Safety">here</a>.
-				</div>
+					<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.
+					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.
+					It has controlled access, biohazard signs and waste disposal bins, and an autoclave. Other safety features are demonstrated below.<br /></p>
+					<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.
+					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 <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.
+					Group 1 organisms are defined as “no or low individual and community risk, a microorganism that is unlikely to cause human or animal disease.”
+					Aseptic technique is used whenever working with these organisms and any contaminated wastes are sterilized by autoclaving or destroyed by commercial pyrolysis.
+					All laboratory chemicals are stored, handled and used as recommended by the manufacturer,
+					and they are disposed of in accordance with national, state, and local regulations and recommendations.
+					The laboratory space and contents are not accessible by unauthorized personnel.
+					All microbial strains, including bacteria rendered antibiotic resistant by transformation, harbor nutritional auxotrophies or other mutations
+					that mitigate the risk of their growing outside of the laboratory or causing disease in healthy humans or animals.
+					Furthermore, none of our  recombinant constructs produce any known contagion or toxin.<p>
+					<h2>BioBrick Safety</h2>
+					<p>None of the BioBrick parts utilized or constructed are known to pose any safety issues.
+					Furthermore, they are well contained by the microbes that harbor them. The risk of unintended transfer to any other organism is minimal.</p>
+					<h2>Biosafety Group</h2>
+					<p>Rose-Hulman does not have a biosafety group, committee or review board other than an Animal Care and Use Committee,
+					which oversees animal research. Safety training and laboratory waste disposal are facilitated by an Environmental Health and Safety Officer, who also serves as a resource for faculty and students.</p>
+					<h2>Future iGEM Safety</h2>
+					<p>The possibility of designing mutually dependent strain/vector systems for routine manipulation and storage of BioBrick parts should be explored.</p>
+				</div>
 			</div>
 		</div>
 	</body>
-</html>

From 2012.igem.org

Latest revision as of 03:57, 4 October 2012

Week 1

Week 2

Week 3

Week 4

Team Break Week

Week 5

Week 6

Week 7

Week 8

Week 9

Week 10

July 31, 2012

August 1, 2012

August 2, 2012

August 3, 2012

August 6, 2012

August 7, 2012

August 8, 2012

August 9, 2012

August 10, 2012

August 11, 2012

August 12, 2012

August 13, 2012

August 14, 2012

August 15, 2012

August 16, 2012

August 17, 2012

August 28, 2012

August 29, 2012

August 30, 2012

August 31, 2012

September 1, 2012

September 2, 2012

September 3, 2012

September 4, 2012

September 5, 2012

September 6, 2012

September 7, 2012

September 8, 2012

September 10, 2012

September 11, 2012

September 12, 2012

September 13, 2012

September 14, 2012

September 15, 2012

September 16, 2012

September 17, 2012

September 18, 2012

September 19, 2012

September 20, 2012

September 22, 2012

September 23, 2012

September 24, 2012

September 25, 2012

September 26, 2012

September 27, 2012

September 28, 2012

September 29, 2012

September 30, 2012

October 1, 2012

October 3, 2012

Strains Used

E. Coli

S. Cerevisiae

Lab Protocols

Researcher, Public, and Environmental Safety

BioBrick Safety

Biosafety Group

Future iGEM Safety