Difference between revisions of "Team:UPMC-Paris/Experiments"

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{{UPMC-Paris}}
 
 
<html>
 
<html>
  
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<div class="column full_size">
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<style>
  
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/********************************* DEFAULT WIKI SETTINGS  ********************************/
  
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<p>Describe the experiments, research and protocols you used in your iGEM project.</p>
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#sideMenu, #top_title {display:none;}
 +
#content { padding:0px; width:1000px; margin-top:-7px; margin-left:0px;}
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</div>
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<div class="column half_size">
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/* styling for the menu items */
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<h5>What should this page contain?</h5>
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.menu_item {
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<ul>
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width:100%;
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<li> Protocols </li>
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margin:-2px 0px 0px -20px;
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<li> Experiments </li>
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padding: 10px 10px; 
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<li>Documentation of the development of your project </li>
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</ul>
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/*STYLING */
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/* styling for the titles */
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 +
.button_click:hover { 
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 +
 
 +
/* IF THE SCREEN IS LESS THAN 1000PX */
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 +
 
 +
 
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/* IF THE SCREEN IS LESS THAN 680PX */
 +
 
 +
@media only screen and (max-width: 680px) {
 +
.collapsable_menu_control { display:block;}
 +
.menu_item {display:none;}
 +
.menu_wrapper { width:100%; height: 15%; position:relative;}
 +
.content_wrapper {width:100%; margin-left:0px;}
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.column.half_size {width:100%; }
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.column img { width: 100%; padding: 5px 0px;}
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.icon {display:block;}
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.highlight {padding:15px 5px;}
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 +
 
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</style>
 +
 
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 +
<!--- THIS IS WHERE THE HTML BEGINS --->
 +
 
 +
 
 +
<!-- This tells the browser that your page is responsive -->
 +
 
 +
<head>
 +
<meta name="viewport" content="width=device-width, initial-scale=1">
 +
</head>
 +
 
 +
 
 +
 
 +
 
 +
<div>
 +
 
 +
 
 +
 
 +
 
 +
<div class="collapsable_menu_control"> MENU ▤ </div>
 +
 
 +
<ul id="accordion" class="accordion">
 +
 
 +
 
 +
<li class="menu_item"> <div class="icon plus"></div> Competent Cells
 +
<ul class="submenu">
 +
<li>  
 +
<a href="">
 +
<div>
 +
<h>Preparation of Bacillus subtilis competent cells</h>
 +
 
 +
<ol><li>Streak out the strain to be made competent on an LB agar plate as a large patch and incubate overnight at 30°C</li>
 +
 
 +
<li>The following morning scrape the cell growth off the plate and use to inoculate fresh, pre-warmed, SpC medium (20 ml) to give an OD600 reading of about 0.5. </li>
 +
 
 +
<li>Incubate the culture at 37°C with vigorous aeration and take periodic OD readings (OD600) to assess cell growth.</li>
 +
 +
<li>When the rate of cell growth is seen to depart from exponential (i.e. no significant change in cell density over 20-30 min) inoculate 200 ml of pre-warmed, SpII medium with 2 ml of stationary-phase culture and continue incubation at 37°C with slower aeration </li>
 +
 
 +
<li>After 90 min incubation, pellet the cells by centrifugation (8,000 g, 5min) at room temperature.</li>
 +
 
 +
<li>Carefully decant the supernatant into a sterile container and save.</li>
 +
 
 +
<li>Gently resuspended the cell pellet in 18 ml of the saved supernatant and add 2 ml of sterile glycerol; mix gently </li>
 +
 
 +
<li>Aliquot the competent cell (0.5 ml) in sterile tubes, freeze rapidly in liquid nitrogen or a dry-iced/ethanol bath or ice/isopropanol bath and store -70.</li></ol>
 +
 
 +
</div></a></li></ul></li>
 +
 
 +
 
 +
<li class="menu_item"> <div class="icon plus"></div> Transformation
 +
<ul class="submenu">
 +
<li>
 +
<a href="">
 +
<div>
 +
<h>Cells Preparation :</h>
 +
 
 +
<ol><li>Thaw competent cells rapidly by immersing frozen tubes in a 37°C water bath</li>
 +
 
 +
<li>Immediately, add one volume of SpII + EGTA to the Thawed cells; mix gently </li>
 +
 
 +
<li>In a sterile test tube add competent cell (0.2~0.5 ml) to the DNA solution (<0.1 ml) and incubate in a roller drum at 37.</li>
 +
 
 +
<li>Dilute the transformed cells as appropriate in T base containing 0.5% glucose and plate immediately onto selective media. </li></ol>
 +
 
 +
<p align="center"><img src="https://static.igem.org/mediawiki/igem.org/d/d3/Transfo_pic_1.png" width="400px"/></p>
 +
</div></a>
 +
<a href=""><div>
 +
<h>Digestion :</h>
 +
<ol><li>Select restriction enzymes to digest your plasmid. Determine an appropriate reaction buffer by reading the instructions for your enzyme. </li>
 +
<p>In a 1.5mL tube combine the following:</p>
 +
<ul><li>DNA </li>
 +
<li>Restriction Enzyme(s)</li>
 +
<li>Buffer </li>
 +
<li>dH2O up to total volume</li> </ul>
 +
<p>Mix gently by pipetting. </p>
 +
 
 +
<li>Incubate tube at appropriate temperature (usually 37°C) for 1 hour. </li>
 +
 
 +
<li>Always follow the manufacturer’s instructions. </li>
 +
 
 +
<li>To visualize the results of your digest, conduct gel electrophoresis</li></ol>
 +
</div></a>
 +
<a href=""><div>
 +
<h>Vector Preparation :</h>
 +
<p>Combine the following in a PCR or Eppendorf tube:</p>
 +
<ul><li>25ng Vector DNA</li>
 +
<li>75ng Insert DNA</li>
 +
<li>Ligase Buffer (1μL/10μL reaction for 10X buffer, and 2μL/10μL reaction for 5X buffer)</li>
 +
<li>0.5-1μL T4 DNA Ligase</li>
 +
<li>H20 to a total of 10μL</li></ul>
 +
<p>Incubate at room temperature for 2hr, or at 16°C overnight (following the manufacturer’s instructions).</p>
 +
 
 +
</div></a></li></ul></li>
 +
 
 +
 
 +
 
 +
<li class="menu_item"> <div class="icon plus"></div> Deletion
 +
<ul class="submenu">
 +
<li>
 +
<a href="">
 +
<div>
 +
<p>Deletion Step 1 :</p>
 +
<img src="aaa" width="500px"/>
 +
<p>Deletion Step 2 :</p>
 +
<img src="aaa" width="500px"/>
 +
<p>Deletion Step 3 :</p>
 +
<img src="aaa" width="500px"/></div></a></li></ul></li>
 +
 
 +
 
 +
<li class="menu_item"> <div class="icon plus"></div> Other
 +
<ul class="submenu">
 +
<li>
 +
<a href="">
 +
<div>
 +
<p>Whatever Step 1 :</p>
 +
<img src="aaa" width="50px"/>
 +
<p>Whatever Step 2 :</p>
 +
<img src="aaa" width="50px"/></div></a>
 +
<a href="">
 +
<div>
 +
<p>Un autre truc Step 1 :</p>
 +
<img src="aaa" width="50px"/>
 +
<p>Un autre truc Step 2 :</p>
 +
<img src="aaa" width="50px"/></div></a></li></ul></li>
  
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</div>
 
  
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<div class="column half_size">
 
−
<h5>Inspiration</h5>
 
−
<ul>
 
−
<li><a href="https://2014.igem.org/Team:Colombia/Protocols">2014 Colombia </a></li>
 
−
<li><a href="https://2014.igem.org/Team:Imperial/Protocols">2014 Imperial </a></li>
 
−
<li><a href="https://2014.igem.org/Team:Caltech/Project/Experiments">2014 Caltech </a></li>
 
 
</ul>
 
</ul>
 +
 
</div>
 
</div>
  
  
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<div class="clear"></div>
 
  
  
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<div class="column half_size">
 
  
 +
<div class="content_wrapper">
  
  
−
</div>
+
 
 +
<h1 id="team_name">  </h1>
 +
<h4 id="page_name">  </h4>
 +
 
 +
 
 +
 
 +
 
 +
 
 +
 
 +
 
 +
 
 +
 
 +
<script>
 +
 
 +
// This is the jquery part of your template.  Try not modify any of this code since it makes your menu work.
 +
 
 +
$(document).ready(function() {
 +
 
 +
$("#HQ_page").attr('id','');
 +
 +
 
 +
if ( wgPageName.substring( 0,  8) == "Template")  {  // if the page is a template it displays the full name in a single line
 +
$("#team_name").html( wgPageName );
 +
}
 +
 
 +
else if (  ( (wgPageName.match(/\//g) || []).length ) == 0 ) {  // if it is the home page , just print the team's name
 +
$("#team_name").html( wgPageName.substring( 5, wgPageName.length ) );
 +
}
 +
 
 +
else {
 +
// this adds the team's name as an h1
 +
$("#team_name").html( wgPageName.substring( 5 , wgPageName.indexOf("/")  ) );
 +
 
 +
// this adds the page's title as an h4
 +
$("#page_name").html (    ( wgPageName.substring( wgPageName.indexOf("/") + 1, wgPageName.length ) ).replace( /\/|_/g , " ")  );
 +
}
 +
 
 +
 
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if( ($(location).attr('href')).indexOf("submit") >= 0  ) {
 +
$(".menu_wrapper").css( { "position": "absolute", "height": "auto"});
 +
$("#wpTextbox1").css( 'margin-top', '440px');
 +
}
 +
 
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 +
 
 +
 
 +
$('#accordion').find('.menu_item').click(function(){
 +
 
 +
//Expand or collapse this panel
 +
submenu = $(this).find('.submenu');
 +
submenu.toggle();
 +
 
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icon = $(this).find('.icon');
 +
 
 +
if ( !$( submenu ).is(':visible') ) {
 +
icon.removeClass("less").addClass("plus");
 +
}
 +
else {
 +
icon.removeClass("plus").addClass("less");
 +
}
 +
 
 +
//Hide the other panels
 +
$(".submenu").not(submenu).hide();
 +
$(".icon").not(icon).removeClass("less").addClass("plus");
 +
});
 +
 
 +
 
 +
$(".collapsable_menu_control").click(function() {
 +
$(".menu_item").toggle();
 +
});
 +
 
 +
$( window ).resize(function() {
 +
$(".menu_item").show();
 +
});
 +
 
 +
 
 +
});
 +
 
 +
 
 +
 
 +
 
 +
</script>
 +
 
  
 
</html>
 
</html>

Revision as of 15:38, 31 July 2016

MENU ▤