Team:Washington/GENERAL PCR PROTOCOL

From 2013.igem.org

(Difference between revisions)
(→')
(→')
 
(2 intermediate revisions not shown)
Line 1: Line 1:
-
<html>
+
{{:Team:Washington/WA_Template}}
-
 
+
-
<style type="text/css">
+
-
.firstHeading {                                                   /* Gets rid of the default iGEM picture*/
+
-
    height:2px;
+
-
    visibility:hidden;
+
-
}
+
-
#catlinks {
+
-
    display: none;
+
-
}
+
-
#content {
+
-
    border-left-width:0px;
+
-
    border-right-width:0px;
+
-
    padding: 0px 5px 0px 5px;
+
-
    width:965px;                                                    /* Content width*/
+
-
}
+
-
#menubar {
+
-
    background-color:  #FFFFFF;                              /* left menu bar color*/
+
-
}
+
-
#menubar ul li a {
+
-
    color: #545353; }                                            /* menu text color*/
+
-
    .right-menu li a {
+
-
    color: red; 
+
-
    background-color: #FFFFFF;                              /* right menu bar color*/
+
-
}
+
-
#p-logo { height 1px; overflow:hidden; display: none;
+
-
}
+
-
#top-section {     
+
-
        background-position: center center;
+
-
        background-repeat: no-repeat;
+
-
        background-color: #FFFFFF;                        /* top header background color*/
+
-
        border-width:0px;
+
-
        height:10px;                        /* top header height (determines how low content will start to appear)*/
+
-
}
+
-
#siteSub {
+
-
display:none;
+
-
}
+
-
#search-controls {                                                /* gets rid of the search bar (it was ugly) */
+
-
display:none;
+
-
margin-top:0px;
+
-
}
+
-
#contentSub {
+
-
display:none;
+
-
}
+
-
 
+
-
/*Now to set heading and body fonts*/
+
-
body {
+
-
background:#000000;
+
-
background-image: url("https://static.igem.org/mediawiki/2012/1/1b/Gradientback.jpg");
+
-
background-position: center;
+
-
background-repeat: repeat-y;
+
-
}               
+
-
h1
+
-
{
+
-
color:#262626;
+
-
text-align:left;
+
-
font-family:"Trebuchet";
+
-
font-size:25px;
+
-
line-height:100%;
+
-
border-bottom: none;
+
-
padding:0px 0px 0px 0px; 
+
-
margin: 0px 0px 0px 0px;
+
-
}
+
-
h2
+
-
{
+
-
color:#262626;
+
-
text-align:left;
+
-
font-family:"Trebuchet";
+
-
font-size:20px;
+
-
line-height: 100%;
+
-
border-bottom: none; 
+
-
padding:0px 0px 0px 0px; 
+
-
margin: 0px 0px 0px 0x;
+
-
}
+
-
h3
+
-
{
+
-
color:#262626;
+
-
text-align:left;
+
-
font-family:"Trebuchet";
+
-
font-size:17px;
+
-
line-height: 100%;
+
-
border-bottom: none; 
+
-
padding:0px 0px 0px 0px; 
+
-
margin: 0px 0px 0px 0x;
+
-
}
+
-
p
+
-
{
+
-
color:#262626;
+
-
text-align: justify;
+
-
font-family:"Arial";
+
-
font-size:14px;
+
-
line-height:100%
+
-
border-bottom: none;
+
-
padding:0px 0px 0px 0px; 
+
-
margin: 0px 0px 0px 0x;
+
-
border:0px
+
-
}
+
-
body {
+
-
color:#808080;
+
-
line-height: 1;
+
-
font-family:"Arial";
+
-
font-size:11px; 
+
-
}
+
-
 
+
-
.whitebox {
+
-
  -moz-box-shadow:  0px 0px 0px #ccc;
+
-
  -webkit-box-shadow:  0px 0px 0px #ccc;
+
-
  box-shadow:  0px 0px 15px #ccc;
+
-
  background-color: #FFFFFF;
+
-
  -moz-border-radius: 1px;
+
-
  border-radius: 15px;
+
-
  margin-top: 20px;
+
-
  padding-top: 20px;
+
-
  padding-left: 20px;
+
-
  padding-right: 20px;
+
-
  padding-bottom: 20px;
+
-
  margin-left: 15px;
+
-
  margin-right: 25px;
+
-
}
+
-
.thumbnail
+
-
{
+
-
margin-bottom:0px;
+
-
padding-bottom:0px;
+
-
position:relative;
+
-
}
+
-
 
+
-
#igemuwmenu, #igemuwmenu ul {
+
-
margin: 0;
+
-
padding: 0;
+
-
list-style: none;
+
-
}
+
-
+
-
#igemuwmenu {
+
-
width: 820px;
+
-
margin: 10px auto;
+
-
border: 1px solid #222;
+
-
background-color: #111;
+
-
background-image: -moz-linear-gradient(#444, #111);
+
-
background-image: -webkit-gradient(linear, left top, left bottom, from(#444), to(#111));
+
-
background-image: -webkit-linear-gradient(#444, #111);
+
-
background-image: -o-linear-gradient(#444, #111);
+
-
background-image: -ms-linear-gradient(#444, #111);
+
-
background-image: linear-gradient(#444, #111);
+
-
-moz-border-radius: 6px;
+
-
-webkit-border-radius: 6px;
+
-
border-radius: 6px;
+
-
-moz-box-shadow: 0 1px 1px #777, 0 1px 0 #666 inset;
+
-
-webkit-box-shadow: 0 1px 1px #777, 0 1px 0 #666 inset;
+
-
box-shadow: 0 1px 1px #777, 0 1px 0 #666 inset;
+
-
                display: inline-block;
+
-
                white-space: nowrap;
+
-
}
+
-
+
-
#igemuwmenu:before,
+
-
#igemuwmenu:after {
+
-
content: "";
+
-
display: table;
+
-
}
+
-
+
-
#igemuwmenu:after {
+
-
clear: both;
+
-
}
+
-
+
-
#igemuwmenu {
+
-
zoom:1;
+
-
}
+
-
+
-
#igemuwmenu li {
+
-
border-right: 1px solid #222;
+
-
-moz-box-shadow: 1px 0 0 #444;
+
-
-webkit-box-shadow: 1px 0 0 #444;
+
-
box-shadow: 1px 0 0 #444;
+
-
position: relative;
+
-
                display: inline-block;
+
-
                white-space: nowrap;
+
-
}
+
-
+
-
#igemuwmenu a {
+
-
padding: 12px 20px;
+
-
color: #999;
+
-
text-transform: uppercase;
+
-
font: bold 11.4px Arial, Helvetica;
+
-
text-decoration: none;
+
-
text-shadow: 0 1px 0 #000;
+
-
                display: inline-block;
+
-
                white-block: nowrap;
+
-
}
+
-
       
+
-
    #igemuwmenu li:hover > a {
+
-
color: #fafafa;
+
-
}
+
-
+
-
*html #igemuwmenu li a:hover { /* IE6 only */
+
-
color: #fafafa;
+
-
}
+
-
+
-
#igemuwmenu ul {
+
-
margin: 20px 0 0 0;
+
-
_margin: 0; /*IE6 only*/
+
-
opacity: 0;
+
-
visibility: hidden;
+
-
position: absolute;
+
-
top: 38px;
+
-
left: 0;
+
-
z-index: 1;   
+
-
background: #444;
+
-
background: -moz-linear-gradient(#444, #111);
+
-
background-image: -webkit-gradient(linear, left top, left bottom, from(#444), to(#111));
+
-
background: -webkit-linear-gradient(#444, #111);   
+
-
background: -o-linear-gradient(#444, #111);
+
-
background: -ms-linear-gradient(#444, #111);
+
-
background: linear-gradient(#444, #111);
+
-
-moz-box-shadow: 0 -1px rgba(255,255,255,.3);
+
-
-webkit-box-shadow: 0 -1px 0 rgba(255,255,255,.3);
+
-
box-shadow: 0 -1px 0 rgba(255,255,255,.3);
+
-
-moz-border-radius: 3px;
+
-
-webkit-border-radius: 3px;
+
-
border-radius: 3px;
+
-
-webkit-transition: all .2s ease-in-out;
+
-
-moz-transition: all .2s ease-in-out;
+
-
-ms-transition: all .2s ease-in-out;
+
-
-o-transition: all .2s ease-in-out;
+
-
transition: all .2s ease-in-out; 
+
-
}
+
-
 
+
-
#igemuwmenu li:hover > ul {
+
-
opacity: 1;
+
-
visibility: visible;
+
-
margin: 0;
+
-
}
+
-
+
-
#igemuwmenu ul ul {
+
-
top: 0;
+
-
left: 150px;
+
-
margin: 0 0 0 20px;
+
-
_margin: 0; /*IE6 only*/
+
-
-moz-box-shadow: -1px 0 0 rgba(255,255,255,.3);
+
-
-webkit-box-shadow: -1px 0 0 rgba(255,255,255,.3);
+
-
box-shadow: -1px 0 0 rgba(255,255,255,.3);
+
-
}
+
-
+
-
#igemuwmenu ul li {
+
-
float: none;
+
-
display: block;
+
-
border: 0;
+
-
_line-height: 0; /*IE6 only*/
+
-
-moz-box-shadow: 0 1px 0 #111, 0 2px 0 #666;
+
-
-webkit-box-shadow: 0 1px 0 #111, 0 2px 0 #666;
+
-
box-shadow: 0 1px 0 #111, 0 2px 0 #666;
+
-
}
+
-
#igemuwmenu ul li.active {
+
-
float: none;
+
-
display: block;
+
-
border: 0;
+
-
_line-height: 0; /*IE6 only*/
+
-
-moz-box-shadow: 0 1px 0 #111, 0 2px 0 #666;
+
-
-webkit-box-shadow: 0 1px 0 #111, 0 2px 0 #666;
+
-
box-shadow: 0 1px 0 #111, 0 2px 0 #666;
+
-
}
+
-
 
+
-
+
-
#igemuwmenu ul li:last-child { 
+
-
-moz-box-shadow: none;
+
-
-webkit-box-shadow: none;
+
-
box-shadow: none;   
+
-
}
+
-
+
-
#igemuwmenu ul a {   
+
-
padding: 10px;
+
-
width: 130px;
+
-
_height: 10px; /*IE6 only*/
+
-
display: block;
+
-
white-space: nowrap;
+
-
float: none;
+
-
text-transform: none;
+
-
}
+
-
     
+
-
        #igemuwmenu ul a.selected {   
+
-
padding: 10px;
+
-
width: 130px;
+
-
_height: 10px; /*IE6 only*/
+
-
display: block;
+
-
white-space: nowrap;
+
-
float: none;
+
-
text-transform: none;
+
-
}
+
-
+
-
#igemuwmenu ul a:hover {
+
-
background-color: #785D84;
+
-
background-image: -moz-linear-gradient(#B6A0BA,  #785D84);
+
-
background-image: -webkit-gradient(linear, left top, left bottom, from(#B6A0BA), to(#785D84));
+
-
background-image: -webkit-linear-gradient(#B6A0BA, #785D84);
+
-
background-image: -o-linear-gradient(#B6A0BA, #785D84);
+
-
background-image: -ms-linear-gradient(#B6A0BA, #785D84);
+
-
background-image: linear-gradient(#B6A0BA, #785D84);
+
-
}
+
-
 
+
-
#igemuwmenu ul li:first-child > a {
+
-
-moz-border-radius: 3px 3px 0 0;
+
-
-webkit-border-radius: 3px 3px 0 0;
+
-
border-radius: 3px 3px 0 0;
+
-
}
+
-
+
-
#igemuwmenu ul li:first-child > a:after {
+
-
content: '';
+
-
position: absolute;
+
-
left: 40px;
+
-
top: -6px;
+
-
border-left: 6px solid transparent;
+
-
border-right: 6px solid transparent;
+
-
border-bottom: 6px solid #444;
+
-
}
+
-
+
-
#igemuwmenu ul ul li:first-child a:after {
+
-
left: -6px;
+
-
top: 50%;
+
-
margin-top: -6px;
+
-
border-left: 0;
+
-
border-bottom: 6px solid transparent;
+
-
border-top: 6px solid transparent;
+
-
border-right: 6px solid #3b3b3b;
+
-
}
+
-
+
-
#igemuwmenu ul li:first-child a:hover:after {
+
-
border-bottom-color: #B6A0BA;
+
-
}
+
-
+
-
#igemuwmenu ul ul li:first-child a:hover:after {
+
-
border-right-color: #0299d3;
+
-
border-bottom-color: transparent;
+
-
}
+
-
+
-
#igemuwmenu ul li:last-child > a {
+
-
-moz-border-radius: 0 0 3px 3px;
+
-
-webkit-border-radius: 0 0 3px 3px;
+
-
border-radius: 0 0 3px 3px;
+
-
}
+
-
+
-
/* Mobile */
+
-
#igemuwmenu-trigger {
+
-
display: none;
+
-
}
+
-
 
+
-
@media screen and (max-width: 600px) {
+
-
 
+
-
/* nav-wrap */
+
-
#igemuwmenu-wrap {
+
-
position: relative;
+
-
}
+
-
 
+
-
#igemuwmenu-wrap * {
+
-
-moz-box-sizing: border-box;
+
-
-webkit-box-sizing: border-box;
+
-
box-sizing: border-box;
+
-
}
+
-
 
+
-
/* menu icon */
+
-
#igemuwmenu-trigger {
+
-
display: block; /* show menu icon */
+
-
height: 40px;
+
-
line-height: 40px;
+
-
cursor: pointer;
+
-
padding: 0 0 0 35px;
+
-
border: 1px solid #222;
+
-
color: #fafafa;
+
-
font-weight: bold;
+
-
background-color: #111;
+
-
background: url(data:image/png;base64,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) no-repeat 10px center, -moz-linear-gradient(#444, #111);
+
-
background: url(data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAABIAAAAPCAMAAADeWG8gAAAAGXRFWHRTb2Z0d2FyZQBBZG9iZSBJbWFnZVJlYWR5ccllPAAAAyBpVFh0WE1MOmNvbS5hZG9iZS54bXAAAAAAADw/eHBhY2tldCBiZWdpbj0i77u/IiBpZD0iVzVNME1wQ2VoaUh6cmVTek5UY3prYzlkIj8+IDx4OnhtcG1ldGEgeG1sbnM6eD0iYWRvYmU6bnM6bWV0YS8iIHg6eG1wdGs9IkFkb2JlIFhNUCBDb3JlIDUuMC1jMDYwIDYxLjEzNDc3NywgMjAxMC8wMi8xMi0xNzozMjowMCAgICAgICAgIj4gPHJkZjpSREYgeG1sbnM6cmRmPSJodHRwOi8vd3d3LnczLm9yZy8xOTk5LzAyLzIyLXJkZi1zeW50YXgtbnMjIj4gPHJkZjpEZXNjcmlwdGlvbiByZGY6YWJvdXQ9IiIgeG1sbnM6eG1wPSJodHRwOi8vbnMuYWRvYmUuY29tL3hhcC8xLjAvIiB4bWxuczp4bXBNTT0iaHR0cDovL25zLmFkb2JlLmNvbS94YXAvMS4wL21tLyIgeG1sbnM6c3RSZWY9Imh0dHA6Ly9ucy5hZG9iZS5jb20veGFwLzEuMC9zVHlwZS9SZXNvdXJjZVJlZiMiIHhtcDpDcmVhdG9yVG9vbD0iQWRvYmUgUGhvdG9zaG9wIENTNSBXaW5kb3dzIiB4bXBNTTpJbnN0YW5jZUlEPSJ4bXAuaWlkOjE2QjAxNjRDOUNEOTExRTE4RTNFRkI1RDQ2MUYxOTQ3IiB4bXBNTTpEb2N1bWVudElEPSJ4bXAuZGlkOjE2QjAxNjREOUNEOTExRTE4RTNFRkI1RDQ2MUYxOTQ3Ij4gPHhtcE1NOkRlcml2ZWRGcm9tIHN0UmVmOmluc3RhbmNlSUQ9InhtcC5paWQ6MTZCMDE2NEE5Q0Q5MTFFMThFM0VGQjVENDYxRjE5NDciIHN0UmVmOmRvY3VtZW50SUQ9InhtcC5kaWQ6MTZCMDE2NEI5Q0Q5MTFFMThFM0VGQjVENDYxRjE5NDciLz4gPC9yZGY6RGVzY3JpcHRpb24+IDwvcmRmOlJERj4gPC94OnhtcG1ldGE+IDw/eHBhY2tldCBlbmQ9InIiPz42AEtnAAAABlBMVEX///////9VfPVsAAAAAnRSTlP/AOW3MEoAAAAWSURBVHjaYmAgFzBiACKFho6NAAEGAD07AG1pn932AAAAAElFTkSuQmCC) no-repeat 10px center, -webkit-linear-gradient(#444, #111);
+
-
background: url(data:image/png;base64,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) no-repeat 10px center, -o-linear-gradient(#444, #111);
+
-
background: url(data:image/png;base64,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) no-repeat 10px center, -ms-linear-gradient(#444, #111);
+
-
background: url(data:image/png;base64,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) no-repeat 10px center, linear-gradient(#444, #111);
+
-
-moz-border-radius: 6px;
+
-
-webkit-border-radius: 6px;
+
-
border-radius: 6px;
+
-
-moz-box-shadow: 0 1px 1px #777, 0 1px 0 #666 inset;
+
-
-webkit-box-shadow: 0 1px 1px #777, 0 1px 0 #666 inset;
+
-
box-shadow: 0 1px 1px #777, 0 1px 0 #666 inset;
+
-
}
+
-
+
-
/* main nav */
+
-
#igemuwmenu {
+
-
margin: 0; padding: 10px;
+
-
position: absolute;
+
-
top: 40px;
+
-
width: 100%;
+
-
z-index: 1;
+
-
background-color: #444;
+
-
display: none;
+
-
-moz-box-shadow: none;
+
-
-webkit-box-shadow: none;
+
-
box-shadow: none;
+
-
}
+
-
 
+
-
#igemuwmenu:after {
+
-
content: '';
+
-
position: absolute;
+
-
left: 25px;
+
-
top: -8px;
+
-
border-left: 8px solid transparent;
+
-
border-right: 8px solid transparent;
+
-
border-bottom: 8px solid #444;
+
-
}
+
-
 
+
-
#igemuwmenu ul {
+
-
position: static;
+
-
visibility: visible;
+
-
opacity: 1;
+
-
margin: 0;
+
-
background: none;
+
-
-moz-box-shadow: none;
+
-
-webkit-box-shadow: none;
+
-
box-shadow: none;
+
-
}
+
-
 
+
-
#igemuwmenu ul ul {
+
-
margin: 0 0 0 20px !important;
+
-
-moz-box-shadow: none;
+
-
-webkit-box-shadow: none;
+
-
box-shadow: none;
+
-
}
+
-
 
+
-
#igemuwmenu li {
+
-
position: static;
+
-
display: block;
+
-
float: none;
+
-
border: 0;
+
-
margin: 5px;
+
-
-moz-box-shadow: none;
+
-
-webkit-box-shadow: none;
+
-
box-shadow: none;
+
-
}
+
-
 
+
-
#igemuwmenu ul li{
+
-
margin-left: 20px;
+
-
-moz-box-shadow: none;
+
-
-webkit-box-shadow: none;
+
-
box-shadow: none;
+
-
}
+
-
 
+
-
#igemuwmenu a{
+
-
display: block;
+
-
float: none;
+
-
padding: 0;
+
-
color: #999;
+
-
}
+
-
 
+
-
#igemuwmenu a:hover{
+
-
color: #fafafa;
+
-
}
+
-
 
+
-
#igemuwmenu ul a{
+
-
padding: 0;
+
-
width: auto;
+
-
}
+
-
 
+
-
#igemuwmenu ul a:hover{
+
-
background: none;
+
-
}
+
-
 
+
-
#igemuwmenu ul li:first-child a:after,
+
-
#igemuwmenu ul ul li:first-child a:after {
+
-
border: 0;
+
-
}
+
-
 
+
-
}
+
-
 
+
-
@media screen and (min-width: 600px) {
+
-
#igemuwmenu {
+
-
display: block !important;
+
-
}
+
-
}
+
-
 
+
-
/* iPad */
+
-
.no-transition {
+
-
-webkit-transition: none;
+
-
-moz-transition: none;
+
-
-ms-transition: none;
+
-
-o-transition: none;
+
-
transition: none;
+
-
opacity: 1;
+
-
visibility: visible;
+
-
display: none; 
+
-
}
+
-
 
+
-
#igemuwmenu li:hover > .no-transition {
+
-
display: block;
+
-
}
+
-
 
+
-
</style>
+
-
 
+
-
<a href="https://2013.igem.org/Team:Washington"><img class="thumbnail"  src='https://static.igem.org/mediawiki/2013/1/1b/Team_Washington2013_banner_final.jpg' width= "965"/></A>
+
-
 
+
-
<a href="http://www.washington.edu/"><img src='https://static.igem.org/mediawiki/2012/2/25/UW_W-Logo_smallRGB.jpg' align="left" width= "57" style="padding-top: 12px; padding-left: 5px;"/></a>
+
-
 
+
-
<a href="https://2012.igem.org/Main_Page"><img src='https://static.igem.org/mediawiki/igem.org/c/c8/IGEM_desat.png' align="right" width= "70"/></a>
+
-
 
+
-
<style type="text/css">
+
-
#content {z-index:4;}
+
-
.expand {background: url('https://static.igem.org/mediawiki/2008/e/ef/Icon-expand.png') no-repeat 95% 50%;}
+
-
.collapse {background: url('https://static.igem.org/mediawiki/2008/c/cd/Icon-collapse.png') no-repeat 95% 50%;}
+
-
.docked {background: #99ccff url("https://static.igem.org/mediawiki/2008/6/62/Ddnavundock.png") no-repeat 50% 50%;}
+
-
.undocked {background: #99ccff url("https://static.igem.org/mediawiki/2008/e/e4/Ddnavdock.png") no-repeat 50% 50%;}
+
-
</style>
+
-
 
+
-
<nav id="igemuwmenu-wrap">   
+
-
 
+
-
<ul id="igemuwmenu">
+
-
 
+
-
<li><a href="https://2013.igem.org/Team:Washington">Home</a>
+
-
<ul>
+
-
<li><a href="https://2013.igem.org/Team:Washington">UW 2013</a></li>
+
-
<li><a href="https://2012.igem.org/Team:Washington">UW 2012</a></li>
+
-
<li><a href="https://2011.igem.org/Team:Washington">UW 2011</a></li>
+
-
<li><a href="https://2010.igem.org/Team:Washington">UW 2010</a></li>
+
-
<li><a href="https://2009.igem.org/Team:Washington">UW 2009</a></li>
+
-
<li><a href="https://2008.igem.org/Team:University_of_Washington">UW 2008</a></li>
+
-
<li><a href="https://2013.igem.org/Main_Page">iGEM Homepage</a></li>
+
-
</ul>
+
-
</li>
+
-
 
+
-
<!--
+
-
<li>
+
-
<a </a> main menu on plastic degradation
+
-
<ul>
+
-
 
+
-
for the drop down of plastic degradation
+
-
format <li><a href="https://2012.igem.org/Team:Washington/Plastics#Background">Background</a></li>
+
-
 
+
-
 
+
-
</ul>
+
-
</li>
+
-
-->
+
-
 
+
-
 
+
-
<li>
+
-
<a href="https://2013.igem.org/Team:Washington/LightSensing">Light Sensing Experiments</a>
+
-
<ul>
+
-
<li><a href="https://2013.igem.org/Team:Washington/LightSensing#Background">Background</a></li>
+
-
<li><a href="https://2013.igem.org/Team:Washington/LightSensing#Cloning">Cloning</a></li>
+
-
<li><a href="https://2013.igem.org/Team:Washington/LightSensing#Systems">Our Systems</a></li>
+
-
<li><a href="https://2013.igem.org/Team:Washington/LightSensing#Methods">Methods</a></li>
+
-
<li><a href="https://2013.igem.org/Team:Washington/LightSensing#Results">Results</a></li>
+
-
<li><a href="https://2013.igem.org/Team:Washington/LightSensing#Sources">Scientific Sources</a></li>
+
-
<li><a href="https://2013.igem.org/Team:Washington/LightSensing#Future">Future Plans</a></li>
+
-
<li><a href="https://2013.igem.org/Team:Washington/LightSensing#Parts">Parts Submitted</a></li>
+
-
</ul>
+
-
</li>
+
-
 
+
-
 
+
-
<li>
+
-
<a href="https://2013.igem.org/Team:Washington/ECOLIGHTTUNE">e.colightTune</a>
+
-
<ul>
+
-
<li><a href="https://2013.igem.org/Team:Washington/ECOLIGHTTUNE#Background">Background</a></li>
+
-
<li><a href="https://2013.igem.org/Team:Washington/ECOLIGHTTUNE#Usage">App</a></li>
+
-
<li><a href="https://2013.igem.org/Team:Washington/ECOLIGHTTUNE#Benefits">Methods</a></li>
+
-
<li><a href="https://2013.igem.org/Team:Washington/ECOLIGHTTUNE#Results">Results Summary</a></li>
+
-
</ul>
+
-
</li>
+
-
 
+
-
<li>
+
-
<a href="https://2013.igem.org/Team:Washington/Outreach">Outreach</a>
+
-
<ul>
+
-
<li><a href="https://2013.igem.org/Team:Washington/Outreach#Activities">Activities</a></li>
+
-
<li><a href="https://2013.igem.org/Team:Washington/Outreach#Pictures">Pictures</a></li>
+
-
</ul>
+
-
</li>
+
-
 
+
-
<li>
+
-
<a href="https://2013.igem.org/Team:Washington/Protocols">Protocols</a>
+
-
<ul>
+
-
<li><a href="https://2013.igem.org/Team:Washington/Protocols#General_Protocols">General</a></li>
+
-
<li><a href="https://2013.igem.org/Team:Washington/Protocols#Cloning_Protocols">Cloning</a></li>
+
-
<li><a href="https://2013.igem.org/Team:Washington/Protocols#Assay">Light assays</a></li>
+
-
</ul>
+
-
</li>
+
-
 
+
-
<li>
+
-
<a href="https://2013.igem.org/Team:Washington/MoreLinks">Links</a>
+
-
<ul>
+
-
<li><a href="https://2013.igem.org/Team:Washington/MoreLinks#Safety">Safety Form</a></li>
+
-
<li><a href="http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2012&group=Washington">Parts Registry</a></li>
+
-
<li><a href="https://2013.igem.org/Team:Washington/MoreLinks#Documentation">Parts Documentation</a></li>
+
-
 
+
-
</ul>
+
-
</li>
+
-
 
+
-
<li style="border-right:0; box-shadow:0px 0 0;">
+
-
<a href="https://2013.igem.org/Team:Washington/Funding">Funding</a>
+
-
</li>
+
-
 
+
-
</li>
+
-
                                                                                               
+
-
</ul>
+
-
</nav>
+
-
 
+
-
<script type="text/javascript" src="http://code.jquery.com/jquery-latest.min.js"></script>
+
-
<script type="text/javascript">
+
-
    $(function() {
+
-
if ($.browser.msie && $.browser.version.substr(0,1)<7)
+
-
{
+
-
$('li').has('ul').mouseover(function(){
+
-
$(this).children('ul').css('visibility','visible');
+
-
}).mouseout(function(){
+
-
$(this).children('ul').css('visibility','hidden');
+
-
})
+
-
}
+
-
 
+
-
/* Mobile */
+
-
$('#igemuwmenu-wrap').prepend('<div id="igemuwmenu-trigger">igemuwmenu</div>');
+
-
$("#igemuwmenu-trigger").on("click", function(){
+
-
$("#menu").slideToggle();
+
-
});
+
-
 
+
-
// iPad
+
-
var isiPad = navigator.userAgent.match(/iPad/i) != null;
+
-
if (isiPad) $('#igemuwmenu ul').addClass('no-transition');     
+
-
    });
+
-
// selection
+
-
 
+
-
</script>
+
-
 
+
-
<script type="text/javascript">
+
-
        $("#content").find("a[href='"+location.href+"']").each(function(){
+
-
        $(this).find("a").addClass("selected")
+
-
        })
+
-
</script>
+
-
 
+
-
<script type="text/javascript">
+
-
 
+
-
  var _gaq = _gaq || [];
+
-
  _gaq.push(['_setAccount', 'UA-26265208-1']);
+
-
  _gaq.push(['_trackPageview']);
+
-
 
+
-
  (function() {
+
-
    var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true;
+
-
    ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js';
+
-
    var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s);
+
-
  })();
+
-
</script>
 
-
</html>
 
-
==''''''==
 
<html>
<html>
 +
<blockquote>
<head>
<head>
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" />

Latest revision as of 04:31, 27 September 2013


Untitled Document

General PCR Protocol

See also: PCR_GoTaq

Resuspending primers:

Protocol: Resuspending PCR primers


Working Stock:

(serves as a backup)

Dilute to 10 times the number on primer vial

10ul fwd primer 90ul DI water

10ul rev primer 90ul DI water


Notes for iGEM 2013 HSB lab:

We are using the GreenTaq polymerase 2X pre-mixed (GoTaq) for PCR.

Use the protocols below for Green Taq polymerase.

Use 50uls for PCR insert amplification. Use 10uls for analytical PCRs (screening).

If PCRing insert use 1ng/ul DNA concentration.


Amplification PCR with Green Taq polymerase:

  1. Setup Amplification PCR Reaction

    1. 1uL Template DNA

    2. 22ul deionized (dI) H2O

    3. 1ul 10uM forward primer (Tm XX C)*

    4. 1ul 10uM reverse Primer (Tm YY C)*

    5. 25ul 2x Green taq (GoTaq)

    6. Total 50ul

  2. Amplification PCR Reaction Program

        1. 95 C for 30s

        2. 95 C for 30s

        3. ZZ* C for 30s (important step)

        4. 72 C for 1min per kb target gene plus 10-15% (important step)

        5. Repeat 2-4 34x

        6. 72 C for 5 to 10min

        7. 4 C for forever (don’t forget to turn the machine off the next day)

  • Tms of all primers must first be calculated after primer design. Annealing temperature (represented by ZZ above) should be 2 degrees C higher than XX or YY, whichever is lower. The temperature doesn’t have to be exact (within 5 degrees or so).


Analytical PCR with Green Taq polymerase:

  1. Setup PCR Reaction

    1. 1uL Template

    2. 2ul dI H2O

    3. 1ul 10uM forward primer (Tm XX C)*

    4. 1ul 10uM reverse Primer (Tm YY C)*

    5. 5ul 2x Green taq (GoTaq)

    6. Total 10uls

  2. PCR Reaction Program

        1. 95 C - 30s

        2. 95 C - 10s

        3. ZZ* C - 10s (important step)

        4. 72 C - 1min per kb target gene plus 10-15% (important step)

        5. Repeat b-d 25x

        6. 72 C - 5 to 10min

        7. 10 or 4 C - forever


Phusion polymerase:

  1. Setup Amplification PCR Reaction

    1. 1uL Template

    2. 1uL 25mM dNTP's

    3. 10uL Phusion HF Buffer

    4. 1.0ul Forward Primer (Tm XX oC)*

    5. 1.0ul Reverse Primer (Tm YY oC)*

    6. 0.5uL Phusion polymerase

    7. 36.5uL diH2O

    8. Total 50uls

  2. Amplification PCR Reaction Program

    1. 98 C - 30s

    2. 98 C - 10s

    3. ZZ* C - 10s (important step)

    4. 72 C - 30s per kb target gene plus 10-15% (important step)

    5. Repeat b-d 29-34x

    6. 72 C - 5 to 10min

    7. 10 or 4 C - forever

  • Tms of all primers must first be calculated after primer design. Annealing temperature (represented by ZZ above) should be 5 C lower than XX or YY, whichever is lower. The temperature doesn’t have to be exact (within 5 degrees or so).

Refer for further details to NEB's online protocol for Phusion:

Protocol for a PCR Reaction using Phusion Hot Start DNA Polymerase

Retrieved from "http://2013.igem.org/Team:Washington/GENERAL_PCR_PROTOCOL"