Team:KAIST Korea/Notebook Labnote

From 2012.igem.org

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(Undo revision 94515 by Horang2k (talk))
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#kaistcontent #inside
 
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#kaistcontent #name
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#kaistcontent ul
#kaistcontent ul
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.accordionButton {
 
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width: 690px;
 
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height:40px;
 
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background-color:transparent;
 
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background-image:url('https://static.igem.org/mediawiki/2012/3/30/Kaist_Protocol_button.png');
 
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cursor: pointer;
 
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}
 
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.accordionButton #protocol {
 
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padding-top:8px;
 
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font-style:italic;
 
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.accordionContent {
 
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width: 690px;
 
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background-color: transparent;
 
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}
 
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.accordionContent #protocolcontent{
 
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background-position:center;
 
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font-family: 'Open Sans', sans-serif;
 
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font-size:10pt;
 
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line-height:120%;
 
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text-align:justify;
 
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padding: 20px 30px 20px 25px;
 
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}
 
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.accordionContent #content #starter
 
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font-family: 'Open Sans', sans-serif;
 
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font-size:14pt;
 
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padding: 20px;
 
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font-size: 1.2em;
 
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}
 
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</style>
</style>
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</style>
 
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</head>
</head>
<body>
<body>
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<div id="main">
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<div id="main">
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<img id="top-img" src="https://static.igem.org/mediawiki/2012/4/4b/Top_image_team_kaist.png"/><!-- 이미지 넣어야대-->
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<img id="top-img" src="https://static.igem.org/mediawiki/2012/6/6d/Back_home.PNG"/>
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<div id="menucontent">
<div id="menucontent">
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<img src="https://static.igem.org/mediawiki/2012/d/df/Part_ico.png"/></a>
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    <img src="https://static.igem.org/mediawiki/2012/4/4d/Note_ico.png"></img>
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<a href="#"><img src="https://static.igem.org/mediawiki/2012/b/b0/Member_ico.png"/></a>
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        <a href="https://2012.igem.org/Team:KAIST_Korea/Notebook_Overview"><img src="https://static.igem.org/mediawiki/2012/e/e3/Overview_ico.png"/></a>
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<a href="#"><img src="https://static.igem.org/mediawiki/2012/c/c3/KAIST_ico.png"/></a>
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        <a href="https://2012.igem.org/Team:KAIST_Korea/Notebook_Protocol"><img src="https://static.igem.org/mediawiki/2012/b/b6/Protocol_ico.png"/></a>
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<a href="#"><img src="https://static.igem.org/mediawiki/2012/e/e6/Gallery_ico.png"/></a>
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        <a href="https://2012.igem.org/Team:KAIST_Korea/Notebook_Labnote"><img src="https://static.igem.org/mediawiki/2012/e/e0/Labnote_ico.png"/></a>
</div>
</div>
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<div id="top-img-description-box"><span id="top-img-description">image description</span></div>
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<div id="top-img-description-box"><span id="top-img-description">aaaa</span></div>
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<img id="horizontal-line" src="https://static.igem.org/mediawiki/2012/7/7f/Horizontal_line_kaist.png"></img>
<img id="horizontal-line" src="https://static.igem.org/mediawiki/2012/7/7f/Horizontal_line_kaist.png"></img>
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<div id="kaistcontent">
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<div id="kaistcontent">
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<div>
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<ul class="tabs">  
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<img id="starter-grad" src="https://static.igem.org/mediawiki/2012/9/95/Starter_gradient_kaist.png"></img>
-
        <li class="active" rel="tab1"> Call of Duty</li>
+
        <h1>Labnote</h1>
-
        <li rel="tab2"> Mortal Combat</li>
+
<span id="sub-title">Please enter sub title</span></br></br></br>
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        <li rel="tab3"> Halo</li>
+
</br></br></br></br></br></br></br></br></br>
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        <li rel="tab4"> Portal</li>
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</div>
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    </ul>
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<div class="tab_container">  
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    <div id="tab1" class="tab_content">
 
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<div id="arc_wrapper">
 
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<div class="accordionButton"><div id="protocol">1. LB agar plate</div></div>
 
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<div class="accordionContent">
 
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<img style="margin:0 auto;float:none;"src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"></img>
 
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<div id="protocolcontent">
 
-
<h3>Materials</h3>
 
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<img id="figure" alt="materials" src="https://static.igem.org/mediawiki/2012/4/46/KAIST_Protocol1_table1.png"></img>
 
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<div style="clear:both;"></div>
 
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</br></br>
 
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<h3>Procedure</h3>
 
-
<ol>
 
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<li>Add 25g LB broth and 15g agar into 1L DDW. </li>
 
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<li>Autoclave</li>
 
-
</ol>
 
-
</div>
 
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<img src="https://static.igem.org/mediawiki/2012/8/85/Protocol_content_bottom.png" id="bot-part"></img>
 
-
</div>
 
-
 
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<div class="accordionButton"><div id="protocol">2. Genomic DNA Purification</div></div>
 
-
<div class="accordionContent">
 
-
<img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/>
 
-
<div id="protocolcontent">
 
-
<h3>Materials</h3>
 
-
 
-
<img id="figure" style="width:632px" alt="materials" src="https://static.igem.org/mediawiki/2012/a/ac/Protocol2_table1.png"></img>
 
-
<div style="clear:both;"></div>
 
-
</br></br>
 
-
 
-
<h3>Procedure</h3></br>
 
-
<h4>Pellet Cells</h4>
 
-
<ol>
 
-
<li>Add 1ml of an overnight culture to a 1.5ml microcentrifuge tube.</li>
 
-
<li>Centrifuge at 13000 rpm for 2.5minutes to pellet the cells.</li>
 
-
<li>Remove the supernatant.</li>
 
-
</ol>
 
-
</br>
 
-
<h4>Lyse Cells</h4>
 
-
<ol start="4">
 
-
<li>Add 1ml of an overnight culture to a 1.5ml microcentrifuge tube.</li>
 
-
<li>Centrifuge at 13000 rpm for 2.5minutes to pellet the cells.</li>
 
-
<li>Remove the supernatant.</li>
 
-
</ol>
 
-
</br>
 
-
<h4>Protein Precipitation</h4>
 
-
<ol start="7">
 
-
<li>Add 200ul of Protein Precipitation Solution to the RNase-treated cell lysate. Vortex vigorously at high speed for 20seconds to mix the Protein Precipitation Solution tieh the cell lysate.</li>
 
-
<li>Incubate the sample on ice for 5minutes.</li>
 
-
<li>Centrifuge at 13,000rpm 3minutes.</li>
 
-
<li>Transfer the supernatant containing the DNA to a clean 1.5ml microcentrifuge tube. </li>
 
-
<span style="padding:none">Note: Some supernatant may remain in the original tube containing the protein pellet. Leave this residual liquid in the tube to avoid contaminating the DNA solution with the precipitated protein.</span>
 
-
<li>Add 600ul isopropanol.</li>
 
-
<li>Gently mix by inversion until the thread-like strands of DNA form a visible mass.</li>
 
-
<li>Centrifuge at 13000rpm for 2minutes.</li>
 
-
<li>Carefully pour off the supernatant. </li>
 
-
<li>Add 600ul of room temperature 70% ethanol and gently invert the tube 5 times to wash the DNA pellet. </li>
 
-
<li>Centrifuge at 13000rpm for 2minutes. Carefully aspirate the ethanol. </li>
 
-
<li>Drain the tube on clean absorbent paper and allow the pellet to air-dry for 10-14minutes. </li>
 
-
<li>Add 50~100 μl TE buffer and rehydrate the DNA by incubating at 65℃ for 20min. Periodically mix the solution by gently tapping the tube. Alternatively, rehydrate the DNA by incubationg the solution overnight at room temperature or at 4℃.</li>
 
-
</ol>
 
-
</br>
 
-
<img id="figure" alt="TE buffer" src="https://static.igem.org/mediawiki/2012/d/df/KAIST_Protocol2_table2.png"></img>
 
-
<div style="clear:both;"></div>
 
-
</div>
 
-
<img src="https://static.igem.org/mediawiki/2012/8/85/Protocol_content_bottom.png" id="bot-part"/>
 
-
</div>
 
-
 
-
 
-
<div class="accordionButton"><div id="protocol">3. Vector transformation</div></div>
 
-
 
-
<div class="accordionContent">
 
-
<img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/>
 
-
<div id="protocolcontent">
 
-
<h3>Procedure</h3></br>
 
-
<h4>1) Restriction enzyme digestion</h4>
 
-
<img id="figure" alt="Restriction enzyme digestion" src="https://static.igem.org/mediawiki/2012/2/27/Kaist_protocol3_table1.png"></img>
 
-
<div style="clear:both;"></div>
 
-
</br>
 
-
<ol>
 
-
<li>Incubate 2hr at 37℃.</li>
 
-
<li>Inactivation 20min at 65℃.</li>
 
-
</ol>
 
-
</br>
 
-
<h4>2) Dephosphorylation (Only for vector)</h4>
 
-
<img id="figure" alt="Dephosphorylation (Only for vector)" src="https://static.igem.org/mediawiki/2012/2/2d/Kaist_protocol3_table2.png"></img>
 
-
<div style="clear:both;"></div>
 
-
</br>
 
-
<ol>
 
-
<li>Incubate 30min at 37℃.</li>
 
-
<li>Inactivation 5min at 70℃.</li>
 
-
</ol>
 
-
</br>
 
-
<h4>3) Ligation</h4>
 
-
<img id="figure" alt="Ligation" src="https://static.igem.org/mediawiki/2012/3/31/Kaist_protocol3_table3.png"></img>
 
-
<div style="clear:both;"></div>
 
-
</br>
 
-
<ol>
 
-
<li>Incubate 16hr at 16℃.</li>
 
-
</ol>
 
-
</br>
 
-
<h4>4) Transformation</h4>
 
-
<ol>
 
-
<li>Add 20ul ligated vector into 100ul of competent cell.</li>
 
-
<li>Incubate ice 5min.</li>
 
-
<li>Heat shock 42℃, 1min 30sec.</li>
 
-
<li>Ice 5min.</li>
 
-
<li>Recovery with 700ul LB at 37℃, 1hr.</li>
 
-
<li>Plating.</li>
 
-
</ol>
 
-
</div>
 
-
<img src="https://static.igem.org/mediawiki/2012/8/85/Protocol_content_bottom.png" id="bot-part"/>
 
-
</div>
 
-
 
-
<div class="accordionButton"><div id="protocol">4. PCR</div></div>
 
-
<div class="accordionContent">
 
-
<img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/>
 
-
<div id="protocolcontent">
 
-
<h3>Procedure</h3>
 
-
<img id="figure" alt="PCR" src="https://static.igem.org/mediawiki/2012/0/02/Kaist_protocol4_table1.png"></img>
 
-
</br></br>
 
-
<h3>Reaction Conditions</h3>
 
-
<ol>
 
-
<li>95℃ 3min</li>
 
-
<li>95℃ 30sec</li>
 
-
<li>54℃ 30sec</li>
 
-
<li>72℃ 1min/kbp</li>
 
-
<li>72℃ 10min</li>
 
-
<li>12℃ ∞</li>
 
-
</ol>
 
-
</div>
 
-
<img src="https://static.igem.org/mediawiki/2012/8/85/Protocol_content_bottom.png" id="bot-part"/>
 
-
</div>
 
-
 
-
<div class="accordionButton"><div id="protocol">5. Gel extraction</div></div>
 
-
<div class="accordionContent">
 
-
<img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/>
 
-
<div id="protocolcontent">
 
-
<h3>Materials</h3>
 
-
<ul>
 
-
<li>MGTM Gel Extraction SV - Macrogen</li>
 
-
</ul>
 
-
</br>
 
-
<h3>Procedure</h3>
 
-
<ol>
 
-
<li>Excise the DNA band of interest using an ethanol-cleaned razor blade.</li>
 
-
<li>Weigh the gel slice in a microcentrifuge tube. Add 3 volumes (ul) of Buffer GB to 1 volume (mg) of gel.</li>
 
-
<li>Incubate at 50°C until the agarose gel is completely melted (5 ~ 10 min).</li>
 
-
<li>Transfer the mixture to a MGTM SV column. Centrifuge for 1 min. Discard the pass-through and re-insert the MGTM SV column into the Collection Tube.</li>
 
-
<li>Add 700 ul of Buffer NW to the MGTM SV column. Centrifuge for 30 sec. Discard the pass-through. Reinsert the MGTM SV column into the Collection Tube.</li>
 
-
<li>Centrifuge for additional 1 min to remove residual wash buffer. Transfer the MGTM SV column to a new 1.5 ml tube.</li>
 
-
<li>Apply 30ul of Buffer EB to the center of the membrane in the MGTM SV column, let stand for 1 min, and centrifuge for 1 min.</li>
 
-
</ol>
 
-
</div>
 
-
<img src="https://static.igem.org/mediawiki/2012/8/85/Protocol_content_bottom.png" id="bot-part"/>
 
-
</div>
 
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    </div><!-- #tab1 -->
 
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    <div id="tab2" class="tab_content">
 
-
 
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      <p><img src="images/mortal combat.jpg"> <br />
 
-
      <strong>
 
-
      Mortal Kombat returns after a lengthy hiatus and puts players
 
-
      back into the Tournament for 2D fighting with gruesome combat.
 
-
      </strong>
 
-
      </p>
 
-
 
-
    </div><!-- #tab2 -->
 
-
    <div id="tab3" class="tab_content">
 
-
 
-
      <p><img src="images/halo.jpg"> <br />
 
-
      <strong>
 
-
      Halo: Reach is the culmination of the superlative combat, sensational
 
-
      multiplayer, and seamless online integration that are the hallmarks
 
-
      of this superb series.
 
-
      </strong>
 
-
      </p>
 
-
 
-
    </div><!-- #tab3 -->
 
-
    <div id="tab4" class="tab_content">
 
-
 
-
      <p><img src="images/portal.jpg"> <br />
 
-
      <strong>
 
-
      Portal 2 is an action shooter game from Valve Software that draws
 
-
      from the original formula of innovative gameplay, story, and music,
 
-
      which earned the original Portal more than 70 industry accolades.
 
-
      </strong>
 
-
      </p>
 
-
 
-
    </div><!-- #tab4 -->
 
-
   
 
-
</div> <!-- .tab_container -->
 
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-
 
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</br>
 
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</br>
 
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</div>
</div>
</div>
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{{:Team:KAIST_Korea/footer}}
{{:Team:KAIST_Korea/footer}}

Revision as of 10:13, 4 September 2012

KAIST Korea 2012 iGEM

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