|
|
| 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_april">
| |
| - | <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">Restriction - Ligation</h1>
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Yoyo Y, Wesley M</li>
| |
| - | <li>Date: April 2, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>J06702 - mCherry</li>
| |
| - | <li>J61002 - RFP Reporter</li>
| |
| - | <li>Promoters: J23113, J23108, J23114, J23100</li>
| |
| - | <li>Cut Promoters at sites E & S</li>
| |
| - | <li>Cut mCherry at sites X & P</li>
| |
| - | <li>Cut Destination Plasmid E1010_pSB1C3 sites E&P</li>
| |
| - | <li class="notebook_nested_start">Promoter Master Mix</li>
| |
| - | <li><ul class="notebook_nested_list">
| |
| - | <li>MilliQ: 66.25uL</li>
| |
| - | <li>BSA: 1.25 uL</li>
| |
| - | <li>Buffer #2: 12.5 uL</li>
| |
| - | <li>DNA: 8 uL</li>
| |
| - | <li>Add 0.5 uL EcoR1 and 0.5 uL Spel to 16 uL MM</li>
| |
| - | </ul>
| |
| - | </li>
| |
| - | </li>
| |
| - | <li class="notebook_nested_start">J06702 Master Mix</li>
| |
| - | <li><ul class="notebook_nested_list">
| |
| - | <li>MilliQ: 66.25 uL</li>
| |
| - | <li>BSA: 1.25 uL</li>
| |
| - | <li>Buffer #2: 12.5 uL</li>
| |
| - | <li>J06702: 40 uL</li>
| |
| - | <li>Add 0.5 uL Xbal and 0.5 uL Pst1 to 24MM</li>
| |
| - | </ul>
| |
| - | </li>
| |
| - | </li>
| |
| - | <li class="notebook_nested_start">pSB1C3 Master Mix</li>
| |
| - | <li><ul class="notebook_nested_list">
| |
| - | <li>MilliQ: 66.25uL</li>
| |
| - | <li>BSA: 1.25 uL</li>
| |
| - | <li>Buffer #2: 12.5 uL</li>
| |
| - | <li>E1010 pSB1C3: 40 uL</li>
| |
| - | <li>Add 0.5 uL EcR1 and 0.5 uL Pst1 to 24MM</li>
| |
| - | </ul>
| |
| - | </li>
| |
| - | </li>
| |
| - | <li>Incubated at 37 degrees Celsuis for 1 hour</li>
| |
| - | <li>Heat killed enzyme for 20mins at 65 degrees Celsius</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">Ligations</h1>
| |
| - | <div class="restrict_width">
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Name: Yoyo Y</li>
| |
| - | <li>Date: April 2, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Tube 1 - J23100_J06702_pSB1C3</li>
| |
| - | <li>Tube 2 - J23108_J06702_pSB1C3</li>
| |
| - | <li>Tube 3 - J23113_J06702_pSB1C3</li>
| |
| - | <li>Tube 4 - J23114_J06702_pSB1C3</li>
| |
| - | <li>Left overnight at 16 degrees Celsuis</li>
| |
| - | <li>Agarose Gel of Digested Parts 1%</li>
| |
| - | <li><img src="images/lethbridgehs2013-4-2-agarose-picture.gif" alt=" " width="400px" /></li>
| |
| - | </ul>
| |
| - | </div>
| |
| - | <ul>
| |
| - | <li>
| |
| - | <table class="table_three table_position">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>Lane #</th>
| |
| - | <th>Contents</th>
| |
| - | <th>Volume</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>1</td>
| |
| - | <td>1 kb DNA Ladder</td>
| |
| - | <td>6µL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>2</td>
| |
| - | <td>E1010 pSB1c3 #4</td>
| |
| - | <td>15µL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>3</td>
| |
| - | <td>E1010 pSB1c3 #3</td>
| |
| - | <td>15µL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>4</td>
| |
| - | <td>E1010 pSB1c3 #2</td>
| |
| - | <td>15µL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>5</td>
| |
| - | <td>E1010 pSB1c3 #1</td>
| |
| - | <td>15µL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>6</td>
| |
| - | <td>J06702 #4</td>
| |
| - | <td>15µL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>7</td>
| |
| - | <td>J06702 #3</td>
| |
| - | <td>15µL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>8</td>
| |
| - | <td>J06702 #2</td>
| |
| - | <td>15µL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>9</td>
| |
| - | <td>J06702 #1</td>
| |
| - | <td>15µL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10</td>
| |
| - | <td>J23100</td>
| |
| - | <td>15µL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>11</td>
| |
| - | <td>J23108</td>
| |
| - | <td>15µL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>12</td>
| |
| - | <td>J23113</td>
| |
| - | <td>15µL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>13</td>
| |
| - | <td>J23114</td>
| |
| - | <td>15µL</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_three">
| |
| - | <h1 class="notebook_entry_header">Transformation</h1>
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Name: Fiona S, Chris I, Yoyo Y</li>
| |
| - | <li>Date: April 3, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Antibiotic: CAM</li>
| |
| - | <li>Part Number and Plasmid: J23100_J06702_pSB1C3, J23108_J06702_pSB1C3, J23113_J06702_pSB1C3, J23114_J06702_pSB1C3</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_four">
| |
| - | <h1 class="notebook_entry_header">Re-Plating Transformation</h1>
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Chris I, Fiona S</li>
| |
| - | <li>Date: April 4, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Part Number and Plasmid: J23100_J06702_pSB1C3, J23108_J06702_pSB1C3, J23113_J06702_pSB1C3, J23114_J06702_pSB1C3</li>
| |
| - | <li>Antibiotic CAM</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">Miniprep of ON Cultures</h1>
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Chris I, Yoyo Y</li>
| |
| - | <li>Date: April 5, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Miniprepped J23108_pSB1A2, J23113_pSB1A2, J23114_pSB1A2, J06702_pSB1AK2, used 50uL of Elution Buffer</li>
| |
| - | </ul>
| |
| - | <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">UV Spectroscopy DNA Concentrations</h1>
| |
| - | <div class="restrict_width">
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Chris I, Yoyo Y, Elaine B</li>
| |
| - | <li>Date: April 5, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | </ul>
| |
| - | </div>
| |
| - | <ul>
| |
| - | <li>
| |
| - | <table class="table_six table_position">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th> </th>
| |
| - | <th>A260</th>
| |
| - | <th>A280</th>
| |
| - | <th colspan="2">A260/A280</th>
| |
| - | <th>DNA Concentration<br /> at 50-fold <br />dilution ug/mL</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>J23108</td>
| |
| - | <td>0.1017</td>
| |
| - | <td>0.0092</td>
| |
| - | <td>11.05435</td>
| |
| - | <td> </td>
| |
| - | <td>254.25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>J23113</td>
| |
| - | <td>0.1289</td>
| |
| - | <td>0.0338</td>
| |
| - | <td>3.813609</td>
| |
| - | <td> </td>
| |
| - | <td>322.25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>J23114</td>
| |
| - | <td>0.135</td>
| |
| - | <td>0.0397</td>
| |
| - | <td>3.400504</td>
| |
| - | <td> </td>
| |
| - | <td>337.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>J06702-1</td>
| |
| - | <td>0.13</td>
| |
| - | <td>0.0373</td>
| |
| - | <td>3.485255</td>
| |
| - | <td> </td>
| |
| - | <td>325</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">Miniprep From Ligation</h1>
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Shikhar M, Krista F, Austin K</li>
| |
| - | <li>Date: April 6, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Miniprepped J23108_J06702_pSB1C3, J23113_J06702_pSB1C3, J23100_J06702_pSB1C3 used 50uL of Elution Buffer</li>
| |
| - | <li>Note: left plates to grow longer, need DNA sequenced.</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">PCR</h1>
| |
| - | <div class="restrict_width">
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Name: Erin K</li>
| |
| - | <li>Date: April 7, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>J23108_J61002, J23113_J61002, J23114_J61002, J06702_pSb1Ak2</li>
| |
| - | <li>Separately added 2uL of appropriate DNA and 0.2uL of Pfu Polymerase to 17.8 uL MM</li>
| |
| - | <li>Agarose Gel confirmation of PCR: Success</li>
| |
| - | </ul>
| |
| - | </div>
| |
| - |
| |
| - | <ul>
| |
| - | <li>
| |
| - | <table class="table_two table_position">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>MM 5x Reagent</th>
| |
| - | <th>Volume(uL)</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tr>
| |
| - | <td>MilliQ H<span class="sub">2</span>O</td>
| |
| - | <td>67</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10X Pfu Buffer with MgSO<span class="sub">4</span></td>
| |
| - | <td>20</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10mM dNTPs</td>
| |
| - | <td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>VF2</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>V4</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tbody>
| |
| - | </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_nine">
| |
| - | <h1 class="notebook_entry_header">Restriction Digestion of PCR Parts</h1>
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Name: Erin K</li>
| |
| - | <li>April 8, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>2 samples of J06702_pSB1AK2: Added 0.5uL XbaI and 0.5uL PstI to 24uL MM</li>
| |
| - | <li>
| |
| - | <table class="table_two_left">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>J06702 MM 2x</th>
| |
| - | <th>Volume (uL)</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>MIlliQ</td>
| |
| - | <td>29.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA</td>
| |
| - | <td>0.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>NEB 10x Buffer 2</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>J06702_pSB1AK2</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | </tbody>
| |
| - | </table>
| |
| - | </li>
| |
| - | <li>2 samples of TAT-E1010_pSB1C3: Added 0.5uL EcoRIand 0.5uL PstI to 24uL MM</li>
| |
| - | <li>
| |
| - | <li>
| |
| - | <table class="table_two_left">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>TAT-E1010_pSB1C3 MM 2x</th>
| |
| - | <th>Volume (uL)</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>MIlliQ</td>
| |
| - | <td>12.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA</td>
| |
| - | <td>0.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>NEB 10x Buffer 2</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TAT-E1010_pSB1C3</td>
| |
| - | <td>30</td>
| |
| - | </tr>
| |
| - | </tbody>
| |
| - | </table>
| |
| - | </li>
| |
| - | </li>
| |
| - | <li>2 samples of each of the 3 promoters: Added 5uL appropriate DNA and 0.5uL EcoRI and 0.5uL SpeI to 16uL MM</li>
| |
| - | <li>
| |
| - | <table class="table_two_left">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>Promoter MM 7x</th>
| |
| - | <th>Volume (uL)</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>MIlliQ</td>
| |
| - | <td>95.75</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA</td>
| |
| - | <td>1.75</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>NEB 10x Buffer 2</td>
| |
| - | <td>17.5</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_ten">
| |
| - | <h1 class="notebook_entry_header">3% Agarose Gel of Digested PCR Parts</h1>
| |
| - | <div class="restrict_width">
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Wesley M, Riley M, Krista F</li>
| |
| - | <li>Date: April 8, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Parts: J23108_J61002, J23113_J61002, J23114_J61002, J06702_pSB1ak2</li>
| |
| - | <li>Results: Unsuccessful</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>Ladder 100bp</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>2</td>
| |
| - | <td>J23108</td>
| |
| - | <td>29</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>3</td>
| |
| - | <td>J23108</td>
| |
| - | <td>29</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>4</td>
| |
| - | <td>Blank</td>
| |
| - | <td>N/A</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>5</td>
| |
| - | <td>J23113</td>
| |
| - | <td>29</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>6</td>
| |
| - | <td>Blank</td>
| |
| - | <td>N/A</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>7</td>
| |
| - | <td>Blank</td>
| |
| - | <td>N/A</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>8</td>
| |
| - | <td>Blank</td>
| |
| - | <td>N/A</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>9</td>
| |
| - | <td>J23113</td>
| |
| - | <td>29</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10</td>
| |
| - | <td>Blank</td>
| |
| - | <td>N/A</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>11</td>
| |
| - | <td>J23114</td>
| |
| - | <td>29</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>12</td>
| |
| - | <td>Blank</td>
| |
| - | <td>N/A</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>13</td>
| |
| - | <td>J23114</td>
| |
| - | <td>29</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_eleven">
| |
| - | <h1 class="notebook_entry_header">Gel Extraction</h1>
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Wesley M, Riley M, Krista F</li>
| |
| - | <li>Date: April 8, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Biobasic Kit: Eluted with 30uL elution buffer</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">Repeat PCR</h1>
| |
| - | <div class="restrict_width">
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Name: Elaine B, Chris I, Fiona S, Wesley M, Riley M</li>
| |
| - | <li>Date: April 9, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>J23108_J61002, J23113_J61002, J23114_J61002, J06702_pSb1Ak2</li>
| |
| - | <li>Separately added 2uL of appropriate DNA and 0.2uL of Pfu Polymerase to 17.8 uL MM</li>\
| |
| - | </ul>
| |
| - | </div>
| |
| - |
| |
| - | <ul>
| |
| - | <li>
| |
| - | <table class="table_two table_position">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>MM 5x Reagent</th>
| |
| - | <th>Volume (μL)</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>MIlliQ</td>
| |
| - | <td>67</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10X Pfu Buffer with MgSO<span class="sub">4</span></td>
| |
| - | <td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10mM dNTPs</td>
| |
| - | <td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>VF2</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>V4</td>
| |
| - | <td>5</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_thirteen">
| |
| - | <h1 class="notebook_entry_header">Picking Cells</h1>
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Chris I, Elaine B, Fiona S, Wesley M, Riley M,</li>
| |
| - | <li>Date: April 9, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Picked: J23113_J61002, J23108_J61002, J23114_J61002, J23100_J61002, J06702_pSB_1AK2</li>
| |
| - | <li>5 mL cultures, 5uL Amp. grown overnight at 37 degrees Celsius</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">Mini Prep</h1>
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Yoyo Y</li>
| |
| - | <li>Date: April 10, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Mini Prep of (x2 of 35uL): J06702_pSB1AK3, J23100_J61002, J23108_J61002, J23113_J61002, J23114_J61002</li>
| |
| - | <li>Mini-prepped with EZ-10 columns</li>
| |
| - | <li>Eluted 35uL</li>
| |
| - | <li>Purified DNA in HS iGEM Kit #1</li>
| |
| - | </ul>
| |
| - | <p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
| |
| - | </div>
| |
| - | </div>
| |
| - |
| |
| - | <div class="notebook_entry">
| |
| - | <div id="entry_fifteen">
| |
| - | <h1 class="notebook_entry_header">Restriction</h1>
| |
| - | <div class="restrict_width">
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Dylan S</li>
| |
| - | <li>Date: April 10, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Add 33uL DNA to 17uL MM</li>
| |
| - | <li>17uL MM to promoter tubes</li>
| |
| - | <li>33uL DNA to promoter tubes</li>
| |
| - | <li>Same procedure for J06702_pSB1AK3</li>
| |
| - | <li>Incubated at 37 degrees Celsius for 2hours</li>
| |
| - | <li>Heat killed at 65 degrees Celsius for 20minutes</li>
| |
| - | </ul>
| |
| - | </div>
| |
| - | <ul>
| |
| - | <li>
| |
| - | <table class="table_two table_position">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>Promoter MM 50μL rxn x4</th>
| |
| - | <th>Volume (μL)</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>MIlliQ</td>
| |
| - | <td>44</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA</td>
| |
| - | <td>20</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>NEB 10 x Buffer 2</td>
| |
| - | <td>20</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_sixteen">
| |
| - | <h1 class="notebook_entry_header">Agarose Gel of PCR 1%</h1>
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Result: Successful</li>
| |
| - | </ul>
| |
| - | <p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
| |
| - | </div>
| |
| - | </div>
| |
| - |
| |
| - | <div class="notebook_entry">
| |
| - | <div id="entry_seventeen">
| |
| - | <h1 class="notebook_entry_header">Miniprep</h1>
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Dylan S, Patrick O</li>
| |
| - | <li>Date: April 11, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Mini prep of (35uL EB): J06702_pSB1AK3, J23100_J61002, J23108_J61002, J23113_J61002, J23114_J61002</li>
| |
| - | <li>Mini-prepped with EZ-10 columns</li>
| |
| - | <li>Eluted 35uL</li>
| |
| - | <li>Purified DNA in HS iGEM Kit #1</li>
| |
| - | </ul>
| |
| - | <p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
| |
| - | </div>
| |
| - | </div>
| |
| - |
| |
| - | <div class="notebook_entry">
| |
| - | <div id="entry_eighteen">
| |
| - | <h1 class="notebook_entry_header">Restriction Digestion</h1>
| |
| - | <div class="restrict_width">
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Wesley M, Yoyo Y</li>
| |
| - | <li>Date: April 12, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Restriction of E1010_pSB1C3 at sites E & P</li>
| |
| - | <li>50uL rxn</li>
| |
| - | <li>Add 30uL DNA to 20uL MM</li>
| |
| - | <li>Added 0.5uL EcroR1 & Pst1</li>
| |
| - | <li>Incubated 2hours at 37 degrees Celsius</li>
| |
| - | <li>Heat killed 20minutes at 65 degrees Celsius</li>
| |
| - | </ul>
| |
| - | </div>
| |
| - | <ul>
| |
| - | <li>
| |
| - | <table class="table_two table_position">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>MM x 2.5</th>
| |
| - | <th>Volume (μL)</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>MIlliQ</td>
| |
| - | <td>22.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10x Buffer 2 (NEB)</td>
| |
| - | <td>12.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA</td>
| |
| - | <td>12.5</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_nineteen">
| |
| - | <h1 class="notebook_entry_header">PCR and Cell Reculturization</h1>
| |
| - | <div class="restrict_width">
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Patrick O, Shikhar M</li>
| |
| - | <li>Date: April 12, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Reculturization of: J06702_pSB1AK3, J23100_J61002, J23108_J61002, J23113_J61002, J23114_J61002</li>
| |
| - | <li>PCR: Master mix created with: Annealing temperature: 55.6 degrees Celsius</li>
| |
| - | </ul>
| |
| - | </div>
| |
| - | <ul>
| |
| - | <li>
| |
| - | <table class="table_four table_position">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th colspan="2">MM for Promoters (x5)</th>
| |
| - | <th colspan="2">Conditions for tube "a"</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>MIlliQ</td>
| |
| - | <td>67 μL</td>
| |
| - | <td>MIlliQ</td>
| |
| - | <td>13.4 μL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10X Pfu Buffer + MgSO<span class="sub">4</span></td>
| |
| - | <td>20 μL</td>
| |
| - | <td>10X Pfu Buffer + MgSO<span class="sub">4</span></td>
| |
| - | <td>4 μL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10mM dNTPs</td>
| |
| - | <td>2 μL</td>
| |
| - | <td>10mM dNTPs</td>
| |
| - | <td>0.4 μL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>VF2</td>
| |
| - | <td>5 μL</td>
| |
| - | <td>VF2</td>
| |
| - | <td>1 μL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>VR</td>
| |
| - | <td>5 μL</td>
| |
| - | <td>VR</td>
| |
| - | <td>1 μL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Pfu polymerase</td>
| |
| - | <td>0.2 μL</td>
| |
| - | <td>Pfu polymerase</td>
| |
| - | <td>0.2 μL</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA part</td>
| |
| - | <td> </td>
| |
| - | <td>2 μL</td>
| |
| - | <td> </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_twenty">
| |
| - | <h1 class="notebook_entry_header">Miniprep</h1>
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Amrinder G, Steven T</li>
| |
| - | <li>Date: April 12, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Mini prep of (35uL EB): J06702_pSB1AK3, J23100_J61002, J23108_J61002 ,J23113_J61002, J23114_J61002</li>
| |
| - | <li>Mini-prepped with EZ-10 columns</li>
| |
| - | <li>Eluted 35uL</li>
| |
| - | <li>Purified DNA in HS iGEM Kit #1</li>
| |
| - | </ul>
| |
| - | <p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
| |
| - | </div>
| |
| - | </div>
| |
| - |
| |
| - | <div class="notebook_entry">
| |
| - | <div id="entry_twenty_one">
| |
| - | <h1 class="notebook_entry_header">Miniprep</h1>
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Fiona S, Katie T</li>
| |
| - | <li>Date: April 14, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Mini prep of (35uL EB): J23100_J61002, J23108_J61002, J23113_J61002, J23114_J61002, J06702_PSB1AK2</li>
| |
| - | <li>Mini-prepped with EZ-10 columns</li>
| |
| - | <li>Eluted 35uL</li>
| |
| - | </ul>
| |
| - | <p class="top_notebook"><a href="#wrapper">Back to Top</a></p>
| |
| - | </div>
| |
| - | </div>
| |
| - |
| |
| - | <div class="notebook_entry">
| |
| - | <div id="entry_twenty_two">
| |
| - | <h1 class="notebook_entry_header">1% Agarose Gel</h1>
| |
| - | <div class="restrict_width">
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Fiona S, Katie T</li>
| |
| - | <li>April 14, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | <li>Parts: J23100_J61002, J23108_J61002, J23113_J61002, J23114_J61002</li>
| |
| - | <li>Gel Extraction</li>
| |
| - | <li>Parts: J23108_J61002, J23113 J61002</li>
| |
| - | <li>Eluted 50 uL</li>
| |
| - | </ul>
| |
| - | </div>
| |
| - | <ul>
| |
| - | <li>
| |
| - | <table class="table_three table_position">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>Lane #</th>
| |
| - | <th>Contents</th>
| |
| - | <th>Volume(µL)</th>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>1</td>
| |
| - | <td>1 µL Dye, 5µL DNA kB ladder</td>
| |
| - | <td>6</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>2</td>
| |
| - | <td>J23100_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>3</td>
| |
| - | <td>J23100_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>4</td>
| |
| - | <td>J23100_J61002</td>
| |
| - | <td>~10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>5</td>
| |
| - | <td>J23108_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>6</td>
| |
| - | <td>J23108_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>7</td>
| |
| - | <td>J23108_J61002</td>
| |
| - | <td>~10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>8</td>
| |
| - | <td>J23113_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>9</td>
| |
| - | <td>J23113_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10</td>
| |
| - | <td>J23113_J61002</td>
| |
| - | <td>~10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>11</td>
| |
| - | <td>J23114_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>12</td>
| |
| - | <td>J23114_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>13</td>
| |
| - | <td>J23114_J61002</td>
| |
| - | <td>~10</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_twenty_six">
| |
| - | <h1 class="notebook_entry_header">Restriction/Digestion</h1>
| |
| - | <div class="restrict_width">
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Master Mix: Yoyo Yao promoters x9</li>
| |
| - | <li>35uL DNA, 14uL MM, 0.5uL Spe1, 0.5uL Pst1</li>
| |
| - | <li>Date: April 15, 2013</li>
| |
| - | </ul>
| |
| - | </div>
| |
| - | <ul>
| |
| - | <li>
| |
| - | <table class="table_two table_position">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>Master Mix</th>
| |
| - | <th>Volume (μL)</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>MIlliQ</td>
| |
| - | <td>36</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA</td>
| |
| - | <td>45</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10x buffer 2 NEB</td>
| |
| - | <td>45</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_twenty_seven">
| |
| - | <h1 class="notebook_entry_header">Restriction/Digestion</h1>
| |
| - | <div class="restrict_width">
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Master Mix: FS, KT ½ promoter ½ insulin/glucose x17</li>
| |
| - | <li>(Promoters) 25uL DNA, 24uL MM, 0.5uL Spe1, 0.5uL Pst1</li>
| |
| - | <li>(Insulin/Glucose) 25uL DNA, 24uL MM, 0.5uL EcoR1, 0.5uL Pst1</li>
| |
| - | <li>Date: April 15, 2013</li>
| |
| - | </ul>
| |
| - | </div>
| |
| - | <ul>
| |
| - | <li>
| |
| - | <table class="table_two table_position">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>Master Mix</th>
| |
| - | <th>Volume (μL)</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>MIlliQ</td>
| |
| - | <td>238</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA</td>
| |
| - | <td>85</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10x buffer 2 NEB</td>
| |
| - | <td>85</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_twenty_three">
| |
| - | <h1 class="notebook_entry_header">1% Agarose Gel of Promoters</h1>
| |
| - | <div class="restrict_width">
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Names: Yoyo Y, Erin K</li>
| |
| - | <li>April 15, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</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>1 KB Ladder</td>
| |
| - | <td>6</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>2</td>
| |
| - | <td>J23100-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>3</td>
| |
| - | <td>J23100-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>4</td>
| |
| - | <td>J23100-1 YYSP</td>
| |
| - | <td><10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>5</td>
| |
| - | <td>J23100-2 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>6</td>
| |
| - | <td>J23100-2 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>7</td>
| |
| - | <td>J23100-2 YYSP</td>
| |
| - | <td><10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>8</td>
| |
| - | <td>J23108-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>9</td>
| |
| - | <td>J23108-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10</td>
| |
| - | <td>J23108-1 YYSP</td>
| |
| - | <td><10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>11</td>
| |
| - | <td>J23108-2 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>12</td>
| |
| - | <td>J23108-2 YYSP</td>
| |
| - | <td>~25</td>
| |
| - | </tr>
| |
| - | </tbody>
| |
| - | </table>
| |
| - | </li>
| |
| - | <li>
| |
| - | <table class="table_three table_position_after_one">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>Lane #</th>
| |
| - | <th>Contents</th>
| |
| - | <th>Volume (μL)</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>1</td>
| |
| - | <td>1 KB Ladder</td>
| |
| - | <td>6</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>2</td>
| |
| - | <td>J23113-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>3</td>
| |
| - | <td>J23113-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>4</td>
| |
| - | <td>J23113-2 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>5</td>
| |
| - | <td>J23113-2 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>6</td>
| |
| - | <td>J23113-2 YYSP</td>
| |
| - | <td><10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>7</td>
| |
| - | <td>J23114-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>8</td>
| |
| - | <td>J23114-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>9</td>
| |
| - | <td>J23114-2 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10</td>
| |
| - | <td>J23114-2 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | </tbody>
| |
| - | </table>
| |
| - | </li>
| |
| - | <li>
| |
| - | <table class="table_three table_position_after_two">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>Lane #</th>
| |
| - | <th>Contents</th>
| |
| - | <th>Volume (μL)</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>1</td>
| |
| - | <td>1 KB Ladder</td>
| |
| - | <td>6</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>2</td>
| |
| - | <td>J23100-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>3</td>
| |
| - | <td>J23100-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>4</td>
| |
| - | <td>J23100-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>5</td>
| |
| - | <td>J23100-2 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>6</td>
| |
| - | <td>J23100-2 YYSP</td>
| |
| - | <td><25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>7</td>
| |
| - | <td>J23108-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>8</td>
| |
| - | <td>J23108-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>9</td>
| |
| - | <td>J23108-1 YYSP</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10</td>
| |
| - | <td>J23108-2 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>11</td>
| |
| - | <td>J23108-2 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | </tbody>
| |
| - | </table>
| |
| - | </li>
| |
| - | <li>
| |
| - | <table class="table_three table_position_after_three">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>Lane #</th>
| |
| - | <th>Contents</th>
| |
| - | <th>Volume (μL)</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>1</td>
| |
| - | <td>1 KB Ladder</td>
| |
| - | <td>6</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>2</td>
| |
| - | <td>J23113-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>3</td>
| |
| - | <td>J23113-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>4</td>
| |
| - | <td>J23113-2 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>5</td>
| |
| - | <td>J23113-2 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>6</td>
| |
| - | <td>J23114-1 YYSP</td>
| |
| - | <td><10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>7</td>
| |
| - | <td>J23114-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>8</td>
| |
| - | <td>J23224-1 YYSP</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>9</td>
| |
| - | <td>J23114-2 YYSP</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10</td>
| |
| - | <td>J23114-2 YYSP</td>
| |
| - | <td>25</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_twenty_four">
| |
| - | <h1 class="notebook_entry_header">Transformation</h1>
| |
| - | <div class="restict_width">
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Name: Dylan S</li>
| |
| - | <li>Date: April 17, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</li>
| |
| - | </ul>
| |
| - | </div>
| |
| - | <ul>
| |
| - | <li>
| |
| - | <table class="table_four table_position">
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th>Tube #</th>
| |
| - | <th>Part #</th>
| |
| - | <th>Plasmid</th>
| |
| - | <th>Volume of Competent Cells(µL)</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>1</td>
| |
| - | <td>J23100+J06702</td>
| |
| - | <td>J61002</td>
| |
| - | <td>100</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>2</td>
| |
| - | <td>J23108+J06702</td>
| |
| - | <td>J61002</td>
| |
| - | <td>100</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>3</td>
| |
| - | <td>J23113+J06702</td>
| |
| - | <td>J61002</td>
| |
| - | <td>50</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>4</td>
| |
| - | <td>J23114+J06702</td>
| |
| - | <td>J61002</td>
| |
| - | <td>50</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>5</td>
| |
| - | <td>J23100 mlc J06702</td>
| |
| - | <td>PSB3C5</td>
| |
| - | <td>50</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>6</td>
| |
| - | <td>J23100 mlc J06702</td>
| |
| - | <td>PSB3C5</td>
| |
| - | <td>50</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>+/- Controls</td>
| |
| - | <td> </td>
| |
| - | <td>PUC19 Amp resistance</td>
| |
| - | <td>50 each</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_twenty_five">
| |
| - | <h1 class="notebook_entry_header">1% Agarose Gel of Promoter Constructs</h1>
| |
| - | <div class="restrict_width">
| |
| - | <ul class="notebook_entry_list">
| |
| - | <li>Name: Chris I</li>
| |
| - | <li>Date: April 30, 2013</li>
| |
| - | <li>Species: E. Coli</li>
| |
| - | <li>Strain: DH5A</li>
| |
| - | <li>Protocol: Ute Kothe</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>1 KB Ladder</td>
| |
| - | <td>6</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>2</td>
| |
| - | <td>J23100_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>3</td>
| |
| - | <td>J23100_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>4</td>
| |
| - | <td>J23100_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>5</td>
| |
| - | <td>J23100_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>6</td>
| |
| - | <td>J23108_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>7</td>
| |
| - | <td>J23108_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>8</td>
| |
| - | <td>J23108_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>9</td>
| |
| - | <td>J23108_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10</td>
| |
| - | <td>J23108_J61002</td>
| |
| - | <td><20</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>11</td>
| |
| - | <td>J23108_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>13</td>
| |
| - | <td>1 KB Ladder</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>12</td>
| |
| - | <td>J23100_J61002</td>
| |
| - | <td><20</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>1</td>
| |
| - | <td>1 KB Ladder</td>
| |
| - | <td>6</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>2</td>
| |
| - | <td>J23113_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>3</td>
| |
| - | <td>J23113_J61002</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>4</td>
| |
| - | <td>J23113_J61002</td>
| |
| - | <td><20</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>5</td>
| |
| - | <td>J6702_pSB1AK2</td>
| |
| - | <td>20</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>6</td>
| |
| - | <td>J6702_pSB1AK2</td>
| |
| - | <td>20</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>7</td>
| |
| - | <td>J6702_pSB1AK2</td>
| |
| - | <td>20</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>8</td>
| |
| - | <td>J6702_pSB1AK2</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>9</td>
| |
| - | <td>J6702_pSB1AK2</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>10</td>
| |
| - | <td>J6702_pSB1AK2</td>
| |
| - | <td>20</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>11</td>
| |
| - | <td>J6702_pSB1AK2</td>
| |
| - | <td>20</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>12</td>
| |
| - | <td>J6702_pSB1AK2</td>
| |
| - | <td>20</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>13</td>
| |
| - | <td>None</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>
| |
| - | </div>
| |
| - | </div>
| |
| - | </div>
| |
| - | </div>
| |
| - | </body>
| |
| - | </html>
| |
"
