Team:Washington/HEAT SHOCK CHEM TRANS

From 2013.igem.org

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<html>
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{{:Team:Washington/WA_Template}}
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<style type="text/css">
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.firstHeading {                                                   /* Gets rid of the default iGEM picture*/
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    height:2px;
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    visibility:hidden;
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}
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#catlinks {
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    display: none;
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}
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#content {
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    border-left-width:0px;
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    border-right-width:0px;
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    padding: 0px 5px 0px 5px;
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    width:965px;                                                    /* Content width*/
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}
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#menubar {
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    background-color:  #FFFFFF;                              /* left menu bar color*/
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}
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#menubar ul li a {
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    color: #545353; }                                            /* menu text color*/
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    .right-menu li a {
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    color: red; 
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    background-color: #FFFFFF;                              /* right menu bar color*/
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}
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#p-logo { height 1px; overflow:hidden; display: none;
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}
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#top-section {     
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        background-position: center center;
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        background-repeat: no-repeat;
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        background-color: #FFFFFF;                        /* top header background color*/
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        border-width:0px;
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        height:10px;                        /* top header height (determines how low content will start to appear)*/
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}
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#siteSub {
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display:none;
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}
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#search-controls {                                                /* gets rid of the search bar (it was ugly) */
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display:none;
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margin-top:0px;
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}
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#contentSub {
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display:none;
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}
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/*Now to set heading and body fonts*/
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body {
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background:#4B0082;
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background-image: url("https://static.igem.org/mediawiki/2012/1/1b/Gradientback.jpg");
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background-position: center;
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background-repeat: repeat-y;
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}               
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h1
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{
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color:#262626;
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text-align:left;
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font-family:"Trebuchet";
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font-size:25px;
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line-height:100%;
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border-bottom: none;
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padding:0px 0px 0px 0px; 
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margin: 0px 0px 0px 0px;
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}
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h2
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{
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color:#262626;
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text-align:left;
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font-family:"Trebuchet";
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font-size:20px;
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line-height: 100%;
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border-bottom: none; 
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padding:0px 0px 0px 0px; 
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margin: 0px 0px 0px 0x;
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}
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h3
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{
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color:#262626;
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text-align:left;
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font-family:"Trebuchet";
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font-size:17px;
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line-height: 100%;
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border-bottom: none; 
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padding:0px 0px 0px 0px; 
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margin: 0px 0px 0px 0x;
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}
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p
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{
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color:#262626;
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text-align: justify;
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font-family:"Arial";
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font-size:14px;
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line-height:100%
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border-bottom: none;
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padding:0px 0px 0px 0px; 
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margin: 0px 0px 0px 0x;
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border:0px
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body {
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color:#808080;
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line-height: 1;
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font-family:"Arial";
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font-size:11px; 
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}
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.whitebox {
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  -moz-box-shadow:  0px 0px 0px #ccc;
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  -webkit-box-shadow:  0px 0px 0px #ccc;
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  box-shadow:  0px 0px 15px #ccc;
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  background-color: #FFFFFF;
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  -moz-border-radius: 1px;
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  border-radius: 15px;
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  margin-top: 20px;
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  padding-top: 20px;
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  padding-left: 20px;
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  padding-right: 20px;
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  padding-bottom: 20px;
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  margin-left: 15px;
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  margin-right: 25px;
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}
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.thumbnail
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{
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margin-bottom:0px;
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padding-bottom:0px;
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position:relative;
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#igemuwmenu, #igemuwmenu ul {
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#igemuwmenu {
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border: 1px solid #222;
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background-color: #111;
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background-image: -moz-linear-gradient(#444, #111);
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background-image: -webkit-gradient(linear, left top, left bottom, from(#444), to(#111));
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background-image: -webkit-linear-gradient(#444, #111);
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background-image: -o-linear-gradient(#444, #111);
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background-image: -ms-linear-gradient(#444, #111);
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background-image: linear-gradient(#444, #111);
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-moz-border-radius: 6px;
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-webkit-border-radius: 6px;
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border-radius: 6px;
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-moz-box-shadow: 0 1px 1px #777, 0 1px 0 #666 inset;
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box-shadow: 0 1px 1px #777, 0 1px 0 #666 inset;
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                display: inline-block;
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#igemuwmenu:before,
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#igemuwmenu:after {
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#igemuwmenu {
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#igemuwmenu li {
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-webkit-box-shadow: 1px 0 0 #444;
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box-shadow: 1px 0 0 #444;
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position: relative;
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                display: inline-block;
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#igemuwmenu a {
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color: #999;
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font: bold 11.4px Arial, Helvetica;
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text-decoration: none;
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    #igemuwmenu li:hover > a {
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color: #fafafa;
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*html #igemuwmenu li a:hover { /* IE6 only */
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color: #fafafa;
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#igemuwmenu ul {
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top: 38px;
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z-index: 1;   
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background: -webkit-linear-gradient(#444, #111);   
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background: -o-linear-gradient(#444, #111);
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-webkit-transition: all .2s ease-in-out;
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-moz-transition: all .2s ease-in-out;
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-ms-transition: all .2s ease-in-out;
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#igemuwmenu ul ul {
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left: 150px;
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#igemuwmenu ul li:last-child { 
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#igemuwmenu ul a {   
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padding: 10px;
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width: 130px;
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        #igemuwmenu ul a.selected {   
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padding: 10px;
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width: 130px;
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display: block;
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white-space: nowrap;
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float: none;
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text-transform: none;
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#igemuwmenu ul a:hover {
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background-image: -moz-linear-gradient(#B6A0BA,  #785D84);
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#igemuwmenu ul li:first-child > a {
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#igemuwmenu ul li:first-child > a:after {
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left: 40px;
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border-left: 6px solid transparent;
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border-bottom: 6px solid #444;
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#igemuwmenu ul ul li:first-child a:after {
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top: 50%;
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#igemuwmenu ul li:first-child a:hover:after {
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#igemuwmenu ul ul li:first-child a:hover:after {
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border-bottom-color: transparent;
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#igemuwmenu ul li:last-child > a {
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-webkit-border-radius: 0 0 3px 3px;
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border-radius: 0 0 3px 3px;
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-
}
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/* Mobile */
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#igemuwmenu-trigger {
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display: none;
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}
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@media screen and (max-width: 600px) {
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/* nav-wrap */
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#igemuwmenu-wrap {
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position: relative;
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#igemuwmenu-wrap * {
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-moz-box-sizing: border-box;
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-webkit-box-sizing: border-box;
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box-sizing: border-box;
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}
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+
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/* menu icon */
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#igemuwmenu-trigger {
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display: block; /* show menu icon */
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height: 40px;
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cursor: pointer;
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padding: 0 0 0 35px;
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font-weight: bold;
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Revision as of 01:28, 25 September 2013


'

Heat shock protocol (approximate time 1hrs)

  1. Remove and thaw (on ice) chemically competent cells, either 1.5ml tube or PCR tube.
    1. 1.5mL tube aliquots of cells contain approx. 205uL cells
    2. Pcr tubes aliquots contain 50uls
  2. Add 1-5µl DNA to 40-50µl of competent cells.
  3. ncubate on ice for 5 minutes (for higher transformation efficiency, do 30 min)
  4. Heat shock cells for 45 seconds at 42°C
  5. Let cells stand on ice for 2 minutes (or longer)
  6. Add 200uL2-1mL of LB and shake for 30 min at 37°C (for higher transformation efficiency, do 1 hr). This is called recovery.
  7. Plate 200uL cells on appropriate antibiotic plate1 and grow O/N in 37°C incubator

Tips
  1. Preheat plates in 37C before for better results (labeling and cell absorption).
  2. If using 300ul pcr tubes, omit shaking, incubate 37C.

NOTE: Do not use XL10 Gold Cells with Chlor Plates

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