Team:Columbia-Cooper-NYC/Miniprep

From 2012.igem.org

(Difference between revisions)
(Plasmid Isolation Protocol)
Line 1: Line 1:
 +
<html>
 +
<head>
 +
<style type="text/css">
 +
body{background-image:url('http://farm9.staticflickr.com/8039/7990439399_1970f791f6_h.jpg');
 +
background-position:top left;
 +
background-repeat:repeat-y
 +
 +
body{background-image:url('http://farm9.staticflickr.com/8173/7989724336_9e5441e3ff_b.jpg');
 +
background-position:top right;
 +
background-repeat:repeat-y
 +
}
 +
 +
 +
 +
</style>
 +
</head>
 +
</html>
 +
 +
 +
 +
<html>
 +
<head>
 +
<body>
 +
<!-- dd menu -->
 +
<script type="text/javascript">
 +
<!--
 +
var timeout        = 500;
 +
var closetimer = 0;
 +
var ddmenuitem      = 0;
 +
 +
// open hidden layer
 +
function mopen(id)
 +
{
 +
// cancel close timer
 +
mcancelclosetime();
 +
 +
// close old layer
 +
if(ddmenuitem) ddmenuitem.style.visibility = 'hidden';
 +
 +
// get new layer and show it
 +
ddmenuitem = document.getElementById(id);
 +
ddmenuitem.style.visibility = 'visible';
 +
 +
}
 +
// close showed layer
 +
function mclose()
 +
{
 +
if(ddmenuitem) ddmenuitem.style.visibility = 'hidden';
 +
}
 +
 +
// go close timer
 +
function mclosetime()
 +
{
 +
closetimer = window.setTimeout(mclose, timeout);
 +
}
 +
 +
// cancel close timer
 +
function mcancelclosetime()
 +
{
 +
if(closetimer)
 +
{
 +
window.clearTimeout(closetimer);
 +
closetimer = null;
 +
}
 +
}
 +
 +
// close layer when click-out
 +
document.onclick = mclose;
 +
// -->
 +
</script>
 +
<style>
 +
#sddm
 +
{ margin: 0;
 +
padding: 0;
 +
      z-index: 30}
 +
 +
#sddm li
 +
{ margin: 0;
 +
padding: 0;
 +
list-style: none;
 +
float: left;
 +
font: bold 11px arial}
 +
 +
#sddm li a
 +
{ display: block;
 +
margin: 0 1px 0 0;
 +
padding: 4px 10px;
 +
width: 85px;
 +
background: #238E23;
 +
color: #FFF;
 +
text-align: center;
 +
text-decoration: none}
 +
 +
#sddm li a:hover
 +
{ background: #B87333}
 +
 +
#sddm div
 +
{ position: absolute;
 +
visibility: hidden;
 +
margin: 0;
 +
padding: 0;
 +
background: #4CC552;
 +
border: 1px solid #5C3317}
 +
 +
#sddm div a
 +
{ position: relative;
 +
display: block;
 +
margin: 0;
 +
padding: 5px 10px;
 +
width: auto;
 +
white-space: nowrap;
 +
text-align: left;
 +
text-decoration: none;
 +
background: #B5EAAA;
 +
color: #5C3317;
 +
font: 11px arial}
 +
 +
#sddm div a:hover
 +
{ background: #49A3FF;
 +
color: #FFF}
 +
</style>
 +
 +
 +
<!-- div class="sample" style="margin-bottom: 15px;height:42px;"><span -->
 +
<ul id="sddm" style="width:1000px;margin:0 auto">
 +
<li><a href="https://2012.igem.org/Team:Columbia-Cooper-NYC" onmouseover="mopen('m1')" onmouseout="mclosetime()">Home</a>
 +
<div id="m1" onmouseover="mcancelclosetime()" onmouseout="mclosetime()">
 +
</div>
 +
</li>
 +
<li><a href="#" onmouseover="mopen('m2')" onmouseout="mclosetime()">Team</a>
 +
<div id="m2" onmouseover="mcancelclosetime()" onmouseout="mclosetime()">
 +
<a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Team">Biographies</a>
 +
<a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Facilitators">Facilitators</a>
 +
<a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Universities">The University(s)</a>
 +
<a href="https://igem.org/Team.cgi?year=2012">Official Profile</a>
 +
</div>
 +
</li>
 +
<li><a href="#" onmouseover="mopen('m3')" onmouseout="mclosetime()">Project</a>
 +
<div id="m3" onmouseover="mcancelclosetime()" onmouseout="mclosetime()">
 +
<a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Overview">Overview</a>
 +
                <a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Data_and_Conclusions">Data and Conclusions</a>
 +
                <a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Future_Directions">Future Directions</a>
 +
                <a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/References">References</a>
 +
</div>
 +
</li>
 +
<li><a href="#" onmouseover="mopen('m4')" onmouseout="mclosetime()">Parts</a>
 +
<div id="m4" onmouseover="mcancelclosetime()" onmouseout="mclosetime()">
 +
<a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Parts_Submitted">Parts Submitted</a>
 +
</div>
 +
</li>
 +
<li><a href="#" onmouseover="mopen('m5')" onmouseout="mclosetime()">Lab Notebook</a>
 +
<div id="m5" onmouseover="mcancelclosetime()" onmouseout="mclosetime()">
 +
<a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Columbia_notebook_2">Genetics</a>
 +
<a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Columbia_notebook_1">Copper Etching</a>
 +
<a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Protocols">Protocols</a>
 +
</div>
 +
</li>
 +
        <li><a href="#" onmouseover="mopen('m6')" onmouseout="mclosetime()">Human Practice</a>
 +
<div id="m6" onmouseover="mcancelclosetime()" onmouseout="mclosetime()">
 +
<a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Outreach">Outreach</a>
 +
<a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Collaborations">Collaboration</a>
 +
</div>
 +
</li>
 +
        <li><a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Safety" onmouseover="mopen('m7')" onmouseout="mclosetime()">Safety</a>
 +
<div id="m7" onmouseover="mcancelclosetime()" onmouseout="mclosetime()">
 +
                <a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Safety">Safety</a>
 +
                <a href="https://2012.igem.org/Security">Security</a>
 +
</div>
 +
</li>
 +
        <li><a href="#" onmouseover="mopen('m8')" onmouseout="mclosetime()">Sponsors</a>
 +
<div id="m8" onmouseover="mcancelclosetime()" onmouseout="mclosetime()">
 +
                <a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Sponsor_Us">Sponsor Us</a>
 +
<a href="https://2012.igem.org/Team:Columbia-Cooper-NYC/Acknowledgements">Acknowledgements</a>
 +
</div>
 +
</li>
 +
        <li><a href="https://2012.igem.org/Main_Page" onmouseover="mopen('m9')" onmouseout="mclosetime()">iGEM</a>
 +
<div id="m9" onmouseover="mcancelclosetime()" onmouseout="mclosetime()">
 +
</div>
 +
</li>
 +
</ul>
 +
<div style="clear:both"></div>
 +
<!-- /span></div -->
 +
<!-- /dd -->
 +
</script>
 +
</body>
 +
</head>
 +
</html>
 +
= Plasmid Isolation Protocol =
= Plasmid Isolation Protocol =
* Following is a plasmid isolation protocol provided by QIAGEN:
* Following is a plasmid isolation protocol provided by QIAGEN:

Revision as of 02:22, 3 October 2012


Plasmid Isolation Protocol

  • Following is a plasmid isolation protocol provided by QIAGEN:
  1. Resuspend pelleted bacterial cells in 250µl Buffer P1 and transfer to a microcentrifuge tube
  2. Add 250µl Buffer P2 and mix thoroughly by inverting the tube 4-6 times
  3. Add 350µl Buffer N3 and mix immediately and thoroughly by inverting the tube 4-6 times
  4. Centrifuge for 10 min. at 13,000 rpm in a table-top microcentrifuge
  5. Apply the supernatant (from step 4) to the QIAprep spin column by decanting
  6. Centrifuge for 30-60 seconds and discard the flow-through
  7. Wash QIAspin column by adding 0.75ml Buffer PE and centrifuging for 30-60 seconds
  8. Discard the flow-through, and centrifuge for an additional 1-3 minutes to remove residual wash buffer
  9. To elute DNA, place the QIAprep column in a clean 1.5ml microcentrifuge tube. Add 50µl water to center of QIAprep spin column, let stand for 1 minute and centrifuge for 1-3 minutes.