Team:Lethbridge Canada/notebook march

From 2013hs.igem.org

(Difference between revisions)
(Blanked the page)
Line 1: Line 1:
-
 
+
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">
 +
<head>
 +
<meta http-equiv="content-type" content="text/html; charset=UTF-8" />
 +
<meta name="viewport" content="width=device-width" />
 +
<title></title>
 +
 +
<link rel="stylesheet" href="bootstrap.css" />
 +
<link rel="stylesheet" href="concept.css" type="text/css" media="screen" />
 +
 +
</head>
 +
<body>
 +
<div id="notebook_march">
 +
<div id="wrapper">
 +
<div id="header">
 +
 +
<a href="index.html">
 +
<div id="logo_all">
 +
<img src="logo.png" alt="logo" />
 +
</div>
 +
<span id="title_first">Lethbridge</span>
 +
<span id="title_second">iGEM Team</span>
 +
</a>
 +
</div>
 +
 +
<div id="content">
 +
 +
<div class="cleafix>">
 +
<div id="navigation">
 +
<ul>
 +
<li class="navigation_project"><a href="#">Project</a>
 +
<ul>
 +
<li></li>
 +
</ul>
 +
</li>
 +
<li class="navigation_parts"><a href="parts.html">Parts</a>
 +
<ul>
 +
<li></li>
 +
</ul>
 +
</li>
 +
<li class="navigation_notebook"><a href="protocols.html">Notebook</a></li>
 +
<li class="navigation_safety"><a href="safety.html">Safety</a></li>
 +
<li class="navigation_outreach"><a href="#">Outreach</a></li>
 +
<li class="navigation_team"><a href="team.html">Team</a></li>
 +
<li class="navigation_attributions"><a href="#">Sponsors</a></li>
 +
</ul>
 +
</div>
 +
</div>
 +
 +
 +
<div id="mainContent">
 +
<div id="notebook_header">
 +
<div id="notebook_links">
 +
<h1 id="notebook_links_header">Notebook Links</h1>
 +
<ul id="notebook_march_links">
 +
<li class="notebook_link_title"><a href="notebook_march.html">March</a></li>
 +
<li><a href="notebook_march.html#entry_one">March 12, 2013: Transformation</a></li>
 +
<li><a href="notebook_march.html#entry_two">March 13, 2013: Picking Cell Colonies (Cultures)</a></li>
 +
<li><a href="notebook_march.html#entry_three">March 13, 2013: Miniprep and Glycerol Stock</a></li>
 +
<li><a href="notebook_march.html#entry_four">March 15, 2013: Restriction Digestion</a></li>
 +
<li><a href="notebook_march.html#entry_five">March 16, 2013: Agarose Gel</a></li>
 +
<li><a href="notebook_march.html#entry_six">March 18, 2013: Agarose Gel</a></li>
 +
<li><a href="notebook_march.html#entry_seven">March 19, 2013: Transformation</a></li>
 +
<li><a href="notebook_march.html#entry_eight">March 20, 2013: Picking Cell Colonies (Cultures)</a></li>
 +
<li><a href="notebook_march.html#entry_nine">March 21, 2013: Miniprep and Glycerol Stock</a></li>
 +
<li><a href="notebook_march.html#entry_ten">March 27, 2013: Transformation</a></li>
 +
<li><a href="notebook_march.html#entry_eleven">March 28, 2013: Transformation</a></li>
 +
<li><a href="notebook_march.html#entry_twelve">March 28, 2013: Picking Cell Colonies (Cultures)</a></li>
 +
<li><a href="notebook_march.html#entry_thirteen">March 29, 2013: Picking Cell Colonies (Cultures)</a></li>
 +
<li><a href="notebook_march.html#entry_fourteen">March 29, 2013: Picking Cells of -80 Glycerol Stock</a></li>
 +
</ul>
 +
 +
<ul id="notebook_april_links">
 +
<li class="notebook_link_title"><a href="notebook_april.html">April</a></li>
 +
<li><a href="notebook_april.html#entry_one">April 2, 2013: Restriction - Ligation</a></li>
 +
<li><a href="notebook_april.html#entry_two">April 2, 2013: Ligation</a></li>
 +
<li><a href="notebook_april.html#entry_three">April 3, 2013: Transformation</a></li>
 +
<li><a href="notebook_april.html#entry_four">April 4, 2013: Re-Plating Transformation</a></li>
 +
<li><a href="notebook_april.html#entry_five">April 5, 2013: Miniprep of ON Cultures</a></li>
 +
<li><a href="notebook_april.html#entry_six">April 5, 2013: UV Spectroscopy DNA Concentrations</a></li>
 +
<li><a href="notebook_april.html#entry_seven">April 6, 2013: Miniprep From Ligations</a></li>
 +
<li><a href="notebook_april.html#entry_eight">April 7, 2013: PCR</a></li>
 +
<li><a href="notebook_april.html#entry_nine">April 8, 2013: Restriction Digestion of PCR Parts</a></li>
 +
<li><a href="notebook_april.html#entry_ten">April 8, 2013: 3% Agarose Gel of Digested PCR Parts</a></li>
 +
<li><a href="notebook_april.html#entry_eleven">April 8, 2013: Gel Extraction</a></li>
 +
<li><a href="notebook_april.html#entry_twelve">April 9, 2013: Repeat PCR</a></li>
 +
<li><a href="notebook_april.html#entry_thirteen">April 9, 2013: Picking Cell Colonies (Cultures)</a></li>
 +
<li><a href="notebook_april.html#entry_fourteen">April 10, 2013: Miniprep</a></li>
 +
<li><a href="notebook_april.html#entry_fifteen">April 10, 2013: Restriction</a></li>
 +
<li><a href="notebook_april.html#entry_sixteen">April 10, 2013: Agarose Gel of PCR 1%</a></li>
 +
<li><a href="notebook_april.html#entry_seventeen">April 11, 2013: Miniprep</a></li>
 +
<li><a href="notebook_april.html#entry_eighteen">April 12, 2013: Restriction Digestion</a></li>
 +
<li><a href="notebook_april.html#entry_nineteen">April 12, 2013: PCR and Cell Reculturization</a></li>
 +
<li><a href="notebook_april.html#entry_twenty">April 12, 2013: Miniprep</a></li>
 +
<li><a href="notebook_april.html#entry_twenty_one">April 14, 2013: Miniprep</a></li>
 +
<li><a href="notebook_april.html#entry_twenty_two">April 14, 2013: 1% Agarose Gel</a></li>
 +
<li><a href="notebook_april.html#entry_twenty_six">April 15, 2013: Restriction/Digestion</a></li>
 +
<li><a href="notebook_april.html#entry_twenty_seven">April 15, 2013: Restriction/Digestion</a></li>
 +
<li><a href="notebook_april.html#entry_twenty_three">April 15, 2013: 1% Agarose Gel of Promoters</a></li>
 +
<li><a href="notebook_april.html#entry_twenty_four">April 17, 2013: Transformation</a></li>
 +
<li><a href="notebook_april.html#entry_twenty_five">April 30, 2013: 1% Agarose Gel of Promoter Constructs</a></li>
 +
</ul>
 +
 +
<ul id="notebook_may_links">
 +
<li class="notebook_link_title"><a href="notebook_may.html">May</a></li>
 +
<li><a href="notebook_may.html#entry_one">May 1, 2013: Ligation</a></li>
 +
<li><a href="notebook_may.html#entry_two">May 4, 2013: Restriction Digest</a></li>
 +
<li><a href="notebook_may.html#entry_three">May 4, 2013: 1% Agarose Gel of Promoter Constructs</a></li>
 +
<li><a href="notebook_may.html#entry_four">May 4, 2013: Gel Extractions</a></li>
 +
<li><a href="notebook_may.html#entry_five">May 5, 2013: Transformation</a></li>
 +
<li><a href="notebook_may.html#entry_six">May 7, 2013: Restriction</a></li>
 +
<li><a href="notebook_may.html#entry_seven">May 7, 2013: Gel Extraction</a></li>
 +
<li><a href="notebook_may.html#entry_eight">May 7, 2013: Gel Extracted on Conformation Gel</a></li>
 +
<li><a href="notebook_may.html#entry_nine">May 8, 2013: Ligations</a></li>
 +
<li><a href="notebook_may.html#entry_ten">May 9, 2013: PCR of Ligation Mixture</a></li>
 +
<li><a href="notebook_may.html#entry_eleven">May 13, 2013: Restriction of PCR Ligations Gel Conformation</a></li>
 +
<li><a href="notebook_may.html#entry_twelve">May 14, 2013: Transformation of Ligations</a></li>
 +
<li><a href="notebook_may.html#entry_thirteen">May 15, 2013: Ligation</a></li>
 +
<li><a href="notebook_may.html#entry_fourteen">May 15, 2013: Picking Cells</a></li>
 +
<li><a href="notebook_may.html#entry_fifteen">May 16, 2013: Picking Cells</a></li>
 +
<li><a href="notebook_may.html#entry_sixteen">May 16, 2013: 10 Fold Miniprep</a></li>
 +
<li><a href="notebook_may.html#entry_seventeen">May 17, 2013: Agarose Gel</a></li>
 +
<li><a href="notebook_may.html#entry_eighteen">May 17, 2013: Miniprep</a></li>
 +
<li><a href="notebook_may.html#entry_nineteen">May 17, 2013: Glycerol Stock</a></li>
 +
</ul>
 +
</div>
 +
</div>
 +
 +
<div id="notebook_body">
 +
<div class="notebook_entry">
 +
<div id="entry_one">
 +
<h1 class="notebook_entry_header">Transformations</h1>
 +
<ul class="notebook_entry_list">
 +
<li >Name: Marie C, Fiona S, Chris I, Katie T, Wesley M</li>
 +
<li >Date: March 12, 2013</li>
 +
<li >Species: E. Coli</li>
 +
<li >Strain: DH5A</li>
 +
<li >Protocol: Ute Kothe</li>
 +
<li >Antibiotic: Kan and Amp</li>
 +
<li >Part Number and Plasmid: R0010 PSB1A2, B0032 PSB1A2, J04650 PSB1AK3, B0031 PSB1A2, B0030 PSB1A2, J04500 PSB1AK3</li>
 +
<li >Results: TBA</li>
 +
</ul>
 +
<p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
 +
</div>
 +
</div>
 +
 +
<div class="notebook_entry">
 +
<div id="entry_two">
 +
<h1 class="notebook_entry_header">Picking Cell Colonies (Cultures)</h1>
 +
<ul class="notebook_entry_list">
 +
<li >Names: Thomas K, Carissa K, Dylan S, Mark H, Brianna C, Wesley M, Austin K, Kieran Mc, Brooke H</li>
 +
<li >Date: March 13, 2013</li>
 +
<li >Species: E. Coli</li>
 +
<li >Strain: DH5A</li>
 +
<li >Protocol: Ute Kothe</li>
 +
<li >Antibiotic: Amp</li>
 +
<li >Part Number and Plasmid: R0010 PSB1A2, B0032 PSB1A2, J04650 PSB1AK3, B0031 PSB1A2, B0030 PSB1A2, J04500 PSB1AK3</li>
 +
<li >Results: TBA</li>
 +
</ul>
 +
<p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
 +
</div>
 +
</div>
 +
 +
<div class="notebook_entry">
 +
<div id="entry_three">
 +
<h1 class="notebook_entry_header">Miniprep and Glycerol Stock</h1>
 +
<ul class="notebook_entry_list">
 +
<li >Names: Mark H, Chris I, Elaine B, Dax L</li>
 +
<li >Date: March 13, 2013</li>
 +
<li >Species: E. Coli</li>
 +
<li >Strain: DH5A</li>
 +
<li >Protocol: Ute Kothe</li>
 +
<li >Miniprepped: 3-B0030, 3-B0031, 3-B0032,
 +
2-J04500 and 3-R0010</li>
 +
<li >Glycerol Stocked B0030 in G1, B0031 in G2, B0032 in G3, J04500 in G4 and R0010 in G5</li>
 +
</ul>
 +
<p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
 +
</div>
 +
</div>
 +
 +
<div class="notebook_entry">
 +
<div id="entry_four">
 +
<h1 class="notebook_entry_header">Restriction Digestion</h1>
 +
<ul class="notebook_entry_list">
 +
<li >Names: Patrick O, Steven T, Amrinder G, Yoyo Y, Wesley M, Orion S</li>
 +
<li >Date: March 15, 2013</li>
 +
<li >Species: E. Coli</li>
 +
<li >Strain: DH5A</li>
 +
<li >Protocol: Ute Kothe use buffer 4 instead of buffer 10</li>
 +
<li >Restricted: 6-B0030, 6-B0031, 6-B0032,
 +
4-J04500 and 6-R0010</li>
 +
<li >Incubate 1 hour at 37˚ C</li>
 +
<li >Stored freezer overnight</li>
 +
</ul>
 +
<p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
 +
</div>
 +
</div>
 +
 +
<div class="notebook_entry">
 +
<div id="entry_five">
 +
<h1 class="notebook_entry_header">Agarose Gel</h1>
 +
<div class="restrict_width">
 +
<ul class="notebook_entry_list">
 +
<li >Name: Patrick O</li>
 +
<li >Date: March 16, 2013</li>
 +
<li >Species: E. Coli</li>
 +
<li >Strain: DH5A</li>
 +
<li >Protocol: Ute Kothe</li>
 +
<li >Ran 15 uL each of 3-B0030, 3-B0031, 3-B0032,
 +
2-J04500 and 3-R0010 through 1% agarose gel</li>
 +
<li >Quantities: 12.5 uL DNA and 2.5 uL of 6x loading dye (15 uL total), 12.5 uL 1kb DNA ladder and 2.5 uL 6x loading dye (15 uL total) note: ladder difficult to see on gel picture, next time less DNA ladder</li>
 +
<ul>
 +
</div>
 +
<table class="table_three table_position">
 +
<thead>
 +
<tr>
 +
<th>Lane #</th>
 +
<th>Contents</th>
 +
<th>Volume(µL)</th>
 +
</tr>
 +
</thead>
 +
<tbody>
 +
<tr>
 +
<td>1</td>
 +
<td>Empty</td>
 +
<td>n/a</td>
 +
</tr>
 +
<tr>
 +
<td>2</td>
 +
<td>1kb DNA Ladder</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>3</td>
 +
<td>BBa_B0030</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>4</td>
 +
<td>BBa_B0030</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>5</td>
 +
<td>BBa_B0030</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>6</td>
 +
<td>BBa_B0031</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>7</td>
 +
<td>BBa_B0031</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>8</td>
 +
<td>BBa_B0031</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>9</td>
 +
<td>BBa_B0032</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>10</td>
 +
<td>BBa_B0032</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>11</td>
 +
<td>BBa_B0032</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>12</td>
 +
<td>BBa_R0010</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>13</td>
 +
<td>BBa_R0010</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>14</td>
 +
<td>BBa_R0010</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>15</td>
 +
<td>BBa_J04500</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>16</td>
 +
<td>BBa_J04500</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>17+</td>
 +
<td>empty</td>
 +
<td>n/a</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
 +
</div>
 +
</div>
 +
 +
<div class="notebook_entry">
 +
<div id="entry_six">
 +
<h1 class="notebook_entry_header">Agarose Gel</h1>
 +
<div class="restrict_width">
 +
<ul class="notebook_entry_list restrict_width">
 +
<li >Names: Fiona S, Katie T, Chris I, Elaine B. </li>
 +
<li >Date: March 18, 2013 </li>
 +
<li >Species: E. Coli</li>
 +
<li >Strain: DH5A</li>
 +
<li >Protocol: Ute Kothe</li>
 +
<li >Ran 15 uL each of 3-B0030, 3-B0031, 3-B0032,
 +
2-J04500 and 3-R0010 through 1% agarose gel</li>
 +
<li >Quantities: 12.5 uL DNA and 2.5 uL of 6x loading dye (15 uL total), 12.5 uL 1kb DNA ladder and 2.5 uL 6x loading dye (15 uL total)</li>
 +
<li ><img src="images/lethbridgehs2013-3-18-agarose-picture.gif" alt"Agrose Gel on March 18, 2013" width="400px" /></li>
 +
</ul>
 +
</div>
 +
<ul>
 +
<li>
 +
<table class="table_three table_position">
 +
<thead>
 +
<tr>
 +
<th>Lane #</th>
 +
<th>Contents</th>
 +
<th>Volume(µL)</th>
 +
</tr>
 +
</thead>
 +
<tbody>
 +
<tr>
 +
<td>1</td>
 +
<td>Empty</td>
 +
<td>n/a</td>
 +
</tr>
 +
<tr>
 +
<td>2</td>
 +
<td>1 kb DNA Ladder</td>
 +
<td>6</td>
 +
</tr>
 +
<tr>
 +
<td>2</td>
 +
<td>1 kb DNA Ladder</td>
 +
<td>6</td>
 +
</tr>
 +
<tr>
 +
<td>3</td>
 +
<td>BBa_B0030</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>4</td>
 +
<td>BBa_B0030</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>5</td>
 +
<td>BBa_B0030</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>6</td>
 +
<td>BBa_B0031</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>7</td>
 +
<td>BBa_B0031</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>8</td>
 +
<td>BBa_B0031</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>9</td>
 +
<td>BBa_B0032</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>10</td>
 +
<td>BBa_B0032</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>11</td>
 +
<td>BBa_B0032</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>12</td>
 +
<td>BBa_R0010</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>13</td>
 +
<td>BBa_R0010</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>14</td>
 +
<td>BBa_R0010</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>15</td>
 +
<td>BBa_J04500</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>16</td>
 +
<td>BBa_J04500</td>
 +
<td>15</td>
 +
</tr>
 +
<tr>
 +
<td>17+</td>
 +
<td>Empty</td>
 +
<td>n/a</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
</li>
 +
</ul>
 +
<p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
 +
</div>
 +
</div>
 +
 +
<div class="notebook_entry">
 +
<div id="entry_seven">
 +
<h1 class="notebook_entry_header">Transformations of J04650</h1>
 +
<ul class="notebook_entry_list">
 +
<li >Names: Fiona S, Chris I.</li>
 +
<li >Date: March 19, 2013 </li>
 +
<li >Species: E. Coli</li>
 +
<li >Strain: DH5A</li>
 +
<li >Protocol: Ute Kothe</li>
 +
<li >Transformed 2 uL of J04650 pSB1AK3 from Spring 2012 Kit Plate 4 C12 into competent cells.</li>
 +
<li >Quantities: 2 uL J04650 pSB1AK3, 200 uL LB media</li>
 +
</ul>
 +
<p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
 +
</div>
 +
</div>
 +
 +
<div class="notebook_entry">
 +
<div id="entry_eight">
 +
<h1 class="notebook_entry_header">Picking Cells of J04650</h1>
 +
<ul class="notebook_entry_list">
 +
<li >Names: Yoyo Y, Wesley M</li>
 +
<li >Date: March 20, 2013</li>
 +
<li >Species: E. Coli</li>
 +
<li >Strain: DH5A</li>
 +
<li >Protocol: Ute Kothe</li>
 +
<li >Picked cells of J04650 from previous day’s transformation</li>
 +
<li >Colonies didn't form properly within either plate, cells were picked with a dab (single colonies)</li>
 +
<li >Quantities: 3x5ml of media</li>
 +
<li >Antibiotic: Ampicillin</li>
 +
<li >Glycerol Stock: R0010 pSB1A2</li>
 +
</ul>
 +
<p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
 +
</div>
 +
</div>
 +
 +
<div class="notebook_entry">
 +
<div id="entry_nine">
 +
<h1 class="notebook_entry_header">Miniprep and Glycerol Stock of R0010</h1>
 +
<ul class="notebook_entry_list">
 +
<li >Name: Mark H</li>
 +
<li >Date: March 21, 2013</li>
 +
<li >Species: E. Coli</li>
 +
<li >Strain: DH5A</li>
 +
<li >Protocol: Ute Kothe</li>
 +
<li >Miniprepped 1-R0010 used 50uL of Elution Buffer</li>
 +
<li >Glycerol Stocked 1-R0010 in G6</li>
 +
</ul>
 +
<p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
 +
</div>
 +
</div>
 +
 +
<div class="notebook_entry">
 +
<div id="entry_ten">
 +
<h1 class="notebook_entry_header">Transformations J04650 and B0015</h1>
 +
<ul class="notebook_entry_list">
 +
<li >Name: Yoyo Y</li>
 +
<li >Date: March 27, 2013</li>
 +
<li >Species: E. Coli</li>
 +
<li >Strain: DH5A</li>
 +
<li >Protocol: Ute Kothe</li>
 +
<li >Transformed 2uL of J04650, and B0015 into 25uL of DH5A cell</li>
 +
<li >Quantities: 2uL of J04650 and 2uL of B0015 added into 25uL of DH5A cells</li>
 +
</ul>
 +
<p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
 +
</div>
 +
</div>
 +
 +
<div class="notebook_entry">
 +
<div id="entry_eleven">
 +
<h1 class="notebook_entry_header">Transformations</h1>
 +
<ul class="notebook_entry_list">
 +
<li >Names: Marie C, Fiona S, Chris I</li>
 +
<li >Date: March 28, 2013</li>
 +
<li >Species: E. Coli</li>
 +
<li >Strain: DH5A</li>
 +
<li >Protocol: Ute Kothe</li>
 +
<li >Transformed 2.5uL of J06702</li>
 +
<li >Plasmid: pSB1A2 </li>
 +
<li >Quantities: 2.5uL of DNA into 100uL of pSB1A2</li>
 +
</ul>
 +
<p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
 +
</div>
 +
</div>
 +
 +
<div class="notebook_entry">
 +
<div id="entry_twelve">
 +
<h1 class="notebook_entry_header">Picking Cells of J04650</h1>
 +
<ul class="notebook_entry_list">
 +
<li >Names: Chris I, Fiona S, Marie C</li>
 +
<li >Date: March 28, 2013</li>
 +
<li >Species: E. Coli</li>
 +
<li >Strain: DH5A</li>
 +
<li >Protocol: Ute Kothe</li>
 +
<li >Picked 3 colonies from March 19, J04650</li>
 +
<li >Picked 1 colony from March 26, J04650</li>
 +
<li >Picked 1 colony from March 27, B0015</li>
 +
<li >5mL cultures with 5uL ampicillin.  </li>
 +
</ul>
 +
<p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
 +
</div>
 +
</div>
 +
 +
<div class="notebook_entry">
 +
<div id="entry_thirteen">
 +
<h1 class="notebook_entry_header">Picking Cells of J06702</h1>
 +
<ul class="notebook_entry_list">
 +
<li >Names: Chris I, Fiona S, Marie C</li>
 +
<li >Date: March 29, 2013</li>
 +
<li >Species: E. Coli</li>
 +
<li >Strain: DH5A</li>
 +
<li >Protocol: Ute Kothe</li>
 +
<li >Picked 2 colonies from March 28, J06702</li>
 +
<li >5mL cultures with 5uL ampicillin</li>
 +
</ul>
 +
<p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
 +
</div>
 +
</div>
 +
 +
<div class="notebook_entry">
 +
<div id="entry_fourteen">
 +
<h1 class="notebook_entry_header">Picking Cells of -80 Glycerol Stock , J61002, J23113, J23108, J23111, J23100</h1>
 +
<ul class="notebook_entry_list">
 +
<li >Names: Chris I, Fiona S, Marie C</li>
 +
<li >Date: March 29, 2013</li>
 +
<li >Species: E. Coli</li>
 +
<li >Strain: DH5A</li>
 +
<li >Protocol: Ute Kothe</li>
 +
<li >Picked, J61002</li>
 +
<li >Picked, J23113</li>
 +
<li >Picked, J23108</li>
 +
<li >Picked, J23114</li>
 +
<li >Picked, J23100 </li>
 +
<li >5mL cultures with 5uL ampicillin</li>
 +
<li >Shaken overnight at 37 degrees Celsius</li>
 +
</ul>
 +
<p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
 +
</div>
 +
</div>
 +
</div>
 +
</div>
 +
</div>
 +
</div>
 +
</div>
 +
</body>
 +
</html>

Revision as of 23:47, 6 June 2013

Transformations

  • Name: Marie C, Fiona S, Chris I, Katie T, Wesley M
  • Date: March 12, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Antibiotic: Kan and Amp
  • Part Number and Plasmid: R0010 PSB1A2, B0032 PSB1A2, J04650 PSB1AK3, B0031 PSB1A2, B0030 PSB1A2, J04500 PSB1AK3
  • Results: TBA

Back to Top

Picking Cell Colonies (Cultures)

  • Names: Thomas K, Carissa K, Dylan S, Mark H, Brianna C, Wesley M, Austin K, Kieran Mc, Brooke H
  • Date: March 13, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Antibiotic: Amp
  • Part Number and Plasmid: R0010 PSB1A2, B0032 PSB1A2, J04650 PSB1AK3, B0031 PSB1A2, B0030 PSB1A2, J04500 PSB1AK3
  • Results: TBA

Back to Top

Miniprep and Glycerol Stock

  • Names: Mark H, Chris I, Elaine B, Dax L
  • Date: March 13, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Miniprepped: 3-B0030, 3-B0031, 3-B0032, 2-J04500 and 3-R0010
  • Glycerol Stocked B0030 in G1, B0031 in G2, B0032 in G3, J04500 in G4 and R0010 in G5

Back to Top

Restriction Digestion

  • Names: Patrick O, Steven T, Amrinder G, Yoyo Y, Wesley M, Orion S
  • Date: March 15, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe use buffer 4 instead of buffer 10
  • Restricted: 6-B0030, 6-B0031, 6-B0032, 4-J04500 and 6-R0010
  • Incubate 1 hour at 37˚ C
  • Stored freezer overnight

Back to Top

Agarose Gel

  • Name: Patrick O
  • Date: March 16, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Ran 15 uL each of 3-B0030, 3-B0031, 3-B0032, 2-J04500 and 3-R0010 through 1% agarose gel
  • Quantities: 12.5 uL DNA and 2.5 uL of 6x loading dye (15 uL total), 12.5 uL 1kb DNA ladder and 2.5 uL 6x loading dye (15 uL total) note: ladder difficult to see on gel picture, next time less DNA ladder
Lane # Contents Volume(µL)
1 Empty n/a
2 1kb DNA Ladder 15
3 BBa_B0030 15
4 BBa_B0030 15
5 BBa_B0030 15
6 BBa_B0031 15
7 BBa_B0031 15
8 BBa_B0031 15
9 BBa_B0032 15
10 BBa_B0032 15
11 BBa_B0032 15
12 BBa_R0010 15
13 BBa_R0010 15
14 BBa_R0010 15
15 BBa_J04500 15
16 BBa_J04500 15
17+ empty n/a

Back to Top

Agarose Gel

  • Names: Fiona S, Katie T, Chris I, Elaine B.
  • Date: March 18, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Ran 15 uL each of 3-B0030, 3-B0031, 3-B0032, 2-J04500 and 3-R0010 through 1% agarose gel
  • Quantities: 12.5 uL DNA and 2.5 uL of 6x loading dye (15 uL total), 12.5 uL 1kb DNA ladder and 2.5 uL 6x loading dye (15 uL total)
  • Lane # Contents Volume(µL)
    1 Empty n/a
    2 1 kb DNA Ladder 6
    2 1 kb DNA Ladder 6
    3 BBa_B0030 15
    4 BBa_B0030 15
    5 BBa_B0030 15
    6 BBa_B0031 15
    7 BBa_B0031 15
    8 BBa_B0031 15
    9 BBa_B0032 15
    10 BBa_B0032 15
    11 BBa_B0032 15
    12 BBa_R0010 15
    13 BBa_R0010 15
    14 BBa_R0010 15
    15 BBa_J04500 15
    16 BBa_J04500 15
    17+ Empty n/a

Back to Top

Transformations of J04650

  • Names: Fiona S, Chris I.
  • Date: March 19, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Transformed 2 uL of J04650 pSB1AK3 from Spring 2012 Kit Plate 4 C12 into competent cells.
  • Quantities: 2 uL J04650 pSB1AK3, 200 uL LB media

Back to Top

Picking Cells of J04650

  • Names: Yoyo Y, Wesley M
  • Date: March 20, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Picked cells of J04650 from previous day’s transformation
  • Colonies didn't form properly within either plate, cells were picked with a dab (single colonies)
  • Quantities: 3x5ml of media
  • Antibiotic: Ampicillin
  • Glycerol Stock: R0010 pSB1A2

Back to Top

Miniprep and Glycerol Stock of R0010

  • Name: Mark H
  • Date: March 21, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Miniprepped 1-R0010 used 50uL of Elution Buffer
  • Glycerol Stocked 1-R0010 in G6

Back to Top

Transformations J04650 and B0015

  • Name: Yoyo Y
  • Date: March 27, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Transformed 2uL of J04650, and B0015 into 25uL of DH5A cell
  • Quantities: 2uL of J04650 and 2uL of B0015 added into 25uL of DH5A cells

Back to Top

Transformations

  • Names: Marie C, Fiona S, Chris I
  • Date: March 28, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Transformed 2.5uL of J06702
  • Plasmid: pSB1A2
  • Quantities: 2.5uL of DNA into 100uL of pSB1A2

Back to Top

Picking Cells of J04650

  • Names: Chris I, Fiona S, Marie C
  • Date: March 28, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Picked 3 colonies from March 19, J04650
  • Picked 1 colony from March 26, J04650
  • Picked 1 colony from March 27, B0015
  • 5mL cultures with 5uL ampicillin.

Back to Top

Picking Cells of J06702

  • Names: Chris I, Fiona S, Marie C
  • Date: March 29, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Picked 2 colonies from March 28, J06702
  • 5mL cultures with 5uL ampicillin

Back to Top

Picking Cells of -80 Glycerol Stock , J61002, J23113, J23108, J23111, J23100

  • Names: Chris I, Fiona S, Marie C
  • Date: March 29, 2013
  • Species: E. Coli
  • Strain: DH5A
  • Protocol: Ute Kothe
  • Picked, J61002
  • Picked, J23113
  • Picked, J23108
  • Picked, J23114
  • Picked, J23100
  • 5mL cultures with 5uL ampicillin
  • Shaken overnight at 37 degrees Celsius

Back to Top

Retrieved from "http://2013hs.igem.org/Team:Lethbridge_Canada/notebook_march"