Difference between revisions of "Team:Peking/Interlab"

Line 1: Line 1:
−
 
 
<!--[if lt IE 8 ]><html class="no-js ie ie7" lang="en"> <![endif]-->
 
<!--[if lt IE 8 ]><html class="no-js ie ie7" lang="en"> <![endif]-->
 
<!--[if IE 8 ]><html class="no-js ie ie8" lang="en"> <![endif]-->
 
<!--[if IE 8 ]><html class="no-js ie ie8" lang="en"> <![endif]-->
Line 8: Line 7:
 
<!--- Basic Page Needs========================================================================= -->
 
<!--- Basic Page Needs========================================================================= -->
 
     <meta charset="utf-8">
 
     <meta charset="utf-8">
−
<title>Standard</title>
+
  <title>Standard</title>
 
     <meta name="viewport" content="width=device-width, initial-scale=1.0, maximum-scale=1.2, user-scalable=yes" />
 
     <meta name="viewport" content="width=device-width, initial-scale=1.0, maximum-scale=1.2, user-scalable=yes" />
 
     <meta name="description" content="Wiki of Peking iGEM 2016" />
 
     <meta name="description" content="Wiki of Peking iGEM 2016" />
Line 14: Line 13:
 
     <meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
 
     <meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
 
<!-- Mobile Specific Metas===================================================================== -->
 
<!-- Mobile Specific Metas===================================================================== -->
−
<meta name="viewport" content="width=device-width, initial-scale=1, maximum-scale=1">
+
  <meta name="viewport" content="width=device-width, initial-scale=1, maximum-scale=1">
 
<!-- Fix  Overwrite the original iGEM style=================================================== -->
 
<!-- Fix  Overwrite the original iGEM style=================================================== -->
 
     <link href="https://2016.igem.org/Template:Peking/css/fix?action=raw&ctype=text/css" rel="stylesheet" />
 
     <link href="https://2016.igem.org/Template:Peking/css/fix?action=raw&ctype=text/css" rel="stylesheet" />
Line 37: Line 36:
 
  <!-- Navigation -->
 
  <!-- Navigation -->
 
     <div id="navigation" class="navbar navbar-fixed-top">
 
     <div id="navigation" class="navbar navbar-fixed-top">
−
<div class="navbar-inner ">
+
  <div class="navbar-inner ">
 
     <div class="container no-padding">
 
     <div class="container no-padding">
−
    <a class="show-menu" data-toggle="collapse" data-target=".nav-collapse"><span class="show-menu-bar"></span>
+
      <a class="show-menu" data-toggle="collapse" data-target=".nav-collapse"><span class="show-menu-bar"></span>
−
</a>
+
    </a>
−
    <div id="logo" style="max-width:170px"><a class="" href="https://2016.igem.org/Team:Peking"></a></div>
+
      <div id="logo" style="max-width:170px"><a class="" href="https://2016.igem.org/Team:Peking"></a></div>
 
          
 
          
 
         <div class="nav-collapse collapse">
 
         <div class="nav-collapse collapse">
−
<ul class="nav">
+
      <ul class="nav">
−
<li class="menu-1"><a class="colapse-menu1" href="https://2016.igem.org/Team:Peking" >Home</a></li>
+
        <li class="menu-1"><a class="colapse-menu1" href="https://2016.igem.org/Team:Peking" >Home</a></li>
−
<li class="dropdown menu-2"><a class="dropdown-toggle" data-toggle="dropdown" href="#">Project</a>
+
        <li class="dropdown menu-2"><a class="dropdown-toggle" data-toggle="dropdown" href="#">Project</a>
 
                   <ul class="dropdown-menu">
 
                   <ul class="dropdown-menu">
 
                       <li><a href="https://2016.igem.org/Team:Peking/Description" >Decription</a></li>
 
                       <li><a href="https://2016.igem.org/Team:Peking/Description" >Decription</a></li>
Line 54: Line 53:
 
                   </ul>
 
                   </ul>
 
                 </li>
 
                 </li>
−
<li class="dropdown menu-3"><a class="dropdown-toggle" data-toggle="dropdown" href="#" >Parts</a>
+
        <li class="dropdown menu-3"><a class="dropdown-toggle" data-toggle="dropdown" href="#" >Parts</a>
 
                   <ul class="dropdown-menu">
 
                   <ul class="dropdown-menu">
 
                       <li><a href="https://2016.igem.org/Team:Peking/Basic_Part" >Basic parts</a></li>
 
                       <li><a href="https://2016.igem.org/Team:Peking/Basic_Part" >Basic parts</a></li>
Line 61: Line 60:
 
                   </ul>
 
                   </ul>
 
                 </li>
 
                 </li>
−
<li class="menu-4"><a class="colapse-menu1" href="https://2016.igem.org/Team:Peking/Human_Practices" >Practices</a></li>
+
        <li class="menu-4"><a class="colapse-menu1" href="https://2016.igem.org/Team:Peking/Human_Practices" >Practices</a></li>
−
<li class="menu-5"><a class="colapse-menu1" href="https://2016.igem.org/Team:Peking/Collaborations" >Collaboration</a></li>
+
        <li class="menu-5"><a class="colapse-menu1" href="https://2016.igem.org/Team:Peking/Collaborations" >Collaboration</a></li>
−
<li class="menu-6"><a class="colapse-menu1" href="https://2016.igem.org/Team:Peking/Model" >Modeling</a></li>
+
        <li class="menu-6"><a class="colapse-menu1" href="https://2016.igem.org/Team:Peking/Model" >Modeling</a></li>
 
                 <li class="dropdown menu-7"><a class="dropdown-toggle" data-toggle="dropdown" href="#" >Lab</a>
 
                 <li class="dropdown menu-7"><a class="dropdown-toggle" data-toggle="dropdown" href="#" >Lab</a>
 
                   <ul class="dropdown-menu">
 
                   <ul class="dropdown-menu">
Line 69: Line 68:
 
                         <li><a class="" href="https://2016.igem.org/Team:Peking/Attributions" >Attribution</a></li>
 
                         <li><a class="" href="https://2016.igem.org/Team:Peking/Attributions" >Attribution</a></li>
 
                         <li><a class="" href="https://2016.igem.org/Team:Peking/Notebook" >Notebook</a></li>
 
                         <li><a class="" href="https://2016.igem.org/Team:Peking/Notebook" >Notebook</a></li>
−
  </ul>
+
          </ul>
 
                 </li>
 
                 </li>
 
                 <li class="dropdown menu-8"><a class="dropdown-toggle" data-toggle="dropdown" href="https://2016.igem.org/Team:Peking/Interlab" >Interlab</a>
 
                 <li class="dropdown menu-8"><a class="dropdown-toggle" data-toggle="dropdown" href="https://2016.igem.org/Team:Peking/Interlab" >Interlab</a>
 
                 </li>
 
                 </li>
−
</div>
+
    </div>
−
</div>
+
  </div>
−
</div>
+
  </div>
−
</div>
+
  </div>
 
     <!--/Navigation -->
 
     <!--/Navigation -->
  
−
<!-- Page Title======================================================================== -->
+
<style type="text/css">
−
  <div id="page-title">
+
−
      <div class="row">
+
−
        <div class="twelve columns centered text-center">
+
−
            <h1>Interlab<span>.</span></h1>
+
−
            <p class="title1" style="text-align:center">This is the special page for team members to learn how to edit.dslkf jadlka gkgjglsg lksdll asd  kdfj asdf adf sadf asdf</p>
+
−
        </div>
+
−
      </div>
+
−
  </div>
+
−
  <!-- Page Title End-->
+
  
−
    
+
#page-wrap {
−
 
+
   width: 25%;
−
<!-- Content=========================================================================== -->
+
  margin: 0px;
−
<div id="page-content" class="row page">
+
  position: relative;
−
<div id="primary" class="twelve columns">
+
}
 +
 
 +
#sidebar {
 +
  width: 25%;
 +
  position: fixed;
 +
  margin-left: 0px;
 +
}
 +
</style>
 +
 
 +
 
 +
<script type="text/javascript">
 +
$(function() {
 +
 
 +
    var $sidebar  = $("#sidebar"),
 +
        $window    = $(window),
 +
        offset    = $sidebar.offset(),
 +
        topPadding = 15;
 +
 
 +
    $window.scroll(function() {
 +
        if ($window.scrollTop() > offset.top) {
 +
            $sidebar.stop().animate({
 +
                marginTop: $window.scrollTop() - offset.top + topPadding
 +
            });
 +
        } else {
 +
            $sidebar.stop().animate({
 +
                marginTop: 0
 +
            });
 +
        }
 +
    });
 +
   
 +
});
 +
</script>
 +
 
 +
 
 +
 
 +
 
 +
 
 +
 
 +
<div id="page-content" class="row page">
 +
    <div id="primary" class="twelve columns">
 
   
 
   
−
<section>
+
      <section>
−
          <div class="row">
+
              <div class="row">
−
           
+
             
 +
             
 +
          <div class="three columns">
 +
            <div id="page-wrap">
 +
<div id="sidebar">
 +
<ul>
 +
  <li>Li Jiamian</li>
 +
    <ul>
 +
        <li><a href="#">Part 1</a></li>
 +
        <li><a href="#">Part 2</a></li>
 +
  <li><a href="#">Wang Yuqing</a></li>
 +
</ul>
 +
</div>
 +
</div>
 +
       
 +
          </div>
 +
         
 +
                 
  
−
         <div class="tweleve columns" style="font-family:arial">
+
         <div class="nine columns">
−
         
+
           
−
             <div>                            
+
             <div>                              
 
                 <h4>Background</h4>
 
                 <h4>Background</h4>
 
                 <p>“All of the 2016 iGEM teams are invited and encouraged to participate in the Third International InterLaboratory Measurement Study in synthetic biology.” Our team took part in this study which aimed to standardize the measurements of fluorescence in different labs. The main task was to quantify expression of GFP in common, comparable or absolute units. In our case, we measured fluorescence using plate reader.
 
                 <p>“All of the 2016 iGEM teams are invited and encouraged to participate in the Third International InterLaboratory Measurement Study in synthetic biology.” Our team took part in this study which aimed to standardize the measurements of fluorescence in different labs. The main task was to quantify expression of GFP in common, comparable or absolute units. In our case, we measured fluorescence using plate reader.
 
</p>
 
</p>
−
              </div>
+
            </div>
 
      
 
      
 
             <div>                                 
 
             <div>                                 
Line 168: Line 214:
 
             </div>
 
             </div>
 
          
 
          
−
       
+
     
 +
 
 
          
 
          
 
          
 
          
 
          
 
          
−
        </div>
+
     
−
 
+
−
 
+
−
 
+
  
  
  
 +
</div>
 
               </div> <!-- row End -->
 
               </div> <!-- row End -->
 +
             
 +
             
 
             </section> <!-- section end -->
 
             </section> <!-- section end -->
 
         </div> <!-- primary end -->
 
         </div> <!-- primary end -->
 
     </div> <!-- page-content End-->
 
     </div> <!-- page-content End-->
 
   </div> <!-- Content End-->
 
   </div> <!-- Content End-->
 +
 +
 +
 +
 +
 +
 +
 +
 +
 +
  
  
Line 234: Line 291:
 
<!--quotations from black: start-->
 
<!--quotations from black: start-->
 
<script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/jquery?action=raw&ctype=text/javascript"></script>
 
<script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/jquery?action=raw&ctype=text/javascript"></script>
−
<script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/jquery_sticky?action=raw&ctype=text/javascript"></script>    
+
<script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/jquery_sticky?action=raw&ctype=text/javascript"></script>      
 
<script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/jquery_easing_1_3_pack?action=raw&ctype=text/javascript"></script>
 
<script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/jquery_easing_1_3_pack?action=raw&ctype=text/javascript"></script>
 
<script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/bootstrap_min?action=raw&ctype=text/javascript"></script>
 
<script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/bootstrap_min?action=raw&ctype=text/javascript"></script>

Revision as of 07:42, 10 September 2016

Standard

Background

“All of the 2016 iGEM teams are invited and encouraged to participate in the Third International InterLaboratory Measurement Study in synthetic biology.” Our team took part in this study which aimed to standardize the measurements of fluorescence in different labs. The main task was to quantify expression of GFP in common, comparable or absolute units. In our case, we measured fluorescence using plate reader.

Design

Fluorescence is widely used as a proxy for promoter activity by expressing fluorescent proteins such as green fluorescent protein (GFP). While this is an indirect measurement, it provides a useful insight into expression levels and has the significant advantage that it can be monitored continuously without disrupting cells.

Fluorescence/OD600 is routinely used to give an adjustment of the relative expression per cell.

We aim to do this using the supplied FITC as a standard reference material. You will measure the fluorescence of your instrument using a dilution series of this reference material to construct a standard curve. We have previously performed this standard curve on our own instrument alongside a standard curve for purified GFP. Using these standard curves alongside your own standard curve for FITC it is thus possible to transform your relative measurements of fluorescence into absolute measurements of GFP molecules.

However, we aim to control for instrument variability, at least to some degree, by measuring a standard scattering solution of a mono-dispersed silica suspension (LUDOX). The objective is to see if a simple, single fixed-point measurement can be used as a ratiometric adjustment to provide greater uniformity in fluorescence/OD600 measurements across sites.

Materials and methods

Used plasmids

• Plasmid DNA (100 pg/uL in 10uL of Buffer EB)

o Test Device 1: J23101.B0034.E0040.B0015 in pSB1C3
o Test Device 2: J23106.B0034.E0040.B0015 in pSB1C3
o Test Device 3: J23117.B0034.E0040.B0015 in pSB1C3
o Positive Control Device: I20270 in pSB1C3 Also located in Kit Plate 3, well 8P
o Negative Control Device: R0040 in pSB1C3 Also located in Kit Plate 2, well 6F

Used strain

• Escherichia coli TOP10

Used material

• FITC Standard: one tube with dried down FITC for creating a FITC standard
• LUDOX: one tube with 30% colloidal silica suspended in 1mL of water
• 1xPBS (phosphate buffered saline)
• Terrific broth (at half strength: 0.5x TB) or can use LB (Luria Bertani) media as an alternative
• Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH)
• 50 ml Falcon tube (or equivalent) or 250 ml shake flask for cell growth
• 1.5 ml eppendorf tubes for sample storage
• Ice bucket with ice
• Pipettes
• 96 well plate

Used machines

• Thermo VARIOSKAN FLASH
• MAPADA UV-3100PC SPECTROPHOTOMETER
• YKKY（FM40）
• AISITE electro-heating standing-temperature cultivator
• HONOUR INCURATOR SHAKER

Used software

• Microsoft Excel