Team:LIKA-CESAR-Brasil/TransformationProtocol

From 2014.igem.org

(Difference between revisions)
(Created page with "{{CSS/Main}} <html lang="pt-br"> <head> <meta charset="utf-8"> <meta http-equiv="X-UA-Compatible" content="IE=edge"> <meta name="viewport" content="width=device-wid...")
 
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<link rel="stylesheet" href="https://maxcdn.bootstrapcdn.com/bootstrap/3.2.0/css/bootstrap.min.css">
<link rel="stylesheet" href="https://maxcdn.bootstrapcdn.com/bootstrap/3.2.0/css/bootstrap.min.css">
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     <link href="https://dl.dropbox.com/s/3g6ibzuen1skq16/lika.css" rel="stylesheet">
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     <style type="text/css">
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body {
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font-size:12px;
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color:#111;
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height:100%;
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background:#ededed;
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/*min-width: 1326px;*/
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}
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.container {
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width:80%;
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background: rgb(255,255,255); /* Old browsers */
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background: -moz-linear-gradient(top,  rgba(255,255,255,1) 0%, rgba(237,237,237,1) 100%); /* FF3.6+ */
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background: -webkit-gradient(linear, left top, left bottom, color-stop(0%,rgba(255,255,255,1)), color-stop(100%,rgba(237,237,237,1))); /* Chrome,Safari4+ */
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background: -webkit-linear-gradient(top,  rgba(255,255,255,1) 0%,rgba(237,237,237,1) 100%); /* Chrome10+,Safari5.1+ */
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background: -o-linear-gradient(top,  rgba(255,255,255,1) 0%,rgba(237,237,237,1) 100%); /* Opera 11.10+ */
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background: -ms-linear-gradient(top,  rgba(255,255,255,1) 0%,rgba(237,237,237,1) 100%); /* IE10+ */
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background: linear-gradient(to bottom,  rgba(255,255,255,1) 0%,rgba(237,237,237,1) 100%); /* W3C */
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filter: progid:DXImageTransform.Microsoft.gradient( startColorstr='#ffffff', endColorstr='#ededed',GradientType=0 ); /* IE6-9 */
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}
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.navbar-brand {
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padding: 5px;
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}
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.col-md-10  {
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width:100%;
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padding: 20px 70px 20px 50px;
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min-height:200px;
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padding-bottom:50px;
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}
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.col-md-4 {
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padding-bottom:70px;
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}
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.navbar-inverse {
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    background-color: #ededed;
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border-color: #ededed;
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}
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.navbar-inverse .navbar-nav > .active > a, .navbar-inverse .navbar-nav > .active > a:hover, .navbar-inverse .navbar-nav > .active > a:focus {
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    color: #006600;
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    background-color: #8cff00;
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.navbar-inverse .navbar-nav > .open > a, .navbar-inverse .navbar-nav > .open > a:hover, .navbar-inverse .navbar-nav > .open > a:focus {
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    color: #006600;
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    background-color: #8cff00;
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}
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.navbar-inverse .navbar-nav > li > a:hover, .navbar-inverse .navbar-nav > li > a:focus {
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    color: #006600;
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    background-color: transparent;
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}
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.navbar-inverse .navbar-nav > li > a:focus {
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    color: #006600;
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    background-color: transparent;
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}
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.nav > li > a {
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    display: block;
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    padding: 25px 15px;
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    position: relative;
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}
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.menu{
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color:black;
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z-index:999;
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position:relative;
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float:right;
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margin:auto;
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}
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.videohome{
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background:#ededed;
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}
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.menu ul{
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margin:0 auto;
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}
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.jumbotron {
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background:url("https://static.igem.org/mediawiki/2014/7/7a/Banner_top.jpg");
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border-top: solid 5px #8cff00;
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border-bottom: solid 5px #8cff00;
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background-size:cover;
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width:80%;
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height:331px;
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margin:auto;
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margin-top:40px;
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}
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.jumbotron02 {
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background:url("https://static.igem.org/mediawiki/2014/5/52/Banner_top02.jpg");
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border-top: solid 5px #8cff00;
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border-bottom: solid 5px #8cff00;
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background-size:cover;
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width:80%;
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height:331px;
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margin:auto;
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margin-top:80px;
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}
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.jumbotronSafety {
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background:url("https://static.igem.org/mediawiki/2014/7/7a/Banner_top.jpg");
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border-top: solid 5px #8cff00;
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border-bottom: solid 5px #8cff00;
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background-size:cover;
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width:80%;
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height:331px;
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margin:auto;
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margin-top:80px;
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}
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.jumbotronParts {
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background:url("https://static.igem.org/mediawiki/2014/b/be/Banner_top04.jpg");
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border-top: solid 5px #8cff00;
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border-bottom: solid 5px #8cff00;
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background-size:cover;
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width:80%;
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height:331px;
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margin:auto;
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margin-top:80px;
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}
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.jumbotronHuman {
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background:url("https://static.igem.org/mediawiki/2014/1/12/Banner_top05.jpg");
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border-top: solid 5px #8cff00;
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border-bottom: solid 5px #8cff00;
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background-size:cover;
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width:80%;
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height:331px;
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margin:auto;
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margin-top:80px;
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}
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.jumbotronProject {
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background:url("https://static.igem.org/mediawiki/2014/b/be/Banner_top04.jpg");
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border-top: solid 5px #8cff00;
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border-bottom: solid 5px #8cff00;
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background-size:cover;
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width:80%;
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height:331px;
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margin:auto;
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margin-top:80px;
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}
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.jumbotronAdvisors {
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background:url("https://static.igem.org/mediawiki/2014/3/3b/Banner_top03.jpg");
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border-top: solid 5px #8cff00;
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border-bottom: solid 5px #8cff00;
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background-size:cover;
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width:80%;
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height:331px;
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margin:auto;
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margin-top:80px;
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}
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.igem-logo{
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    background:url("https://static.igem.org/mediawiki/2014/4/4e/Banner_top_03.png") ;
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}
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/*
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.navbar .container {
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min-width: 1322px;
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    z-index: 9999;
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}
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*/
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h2 {
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font-size: 26px;
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color:#339900;
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width:35%;
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float:right;
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min-height:300px;
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margin-top:10px;
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}
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p {
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font-size: 12px;
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color:#444;
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}
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footer {
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background:#b8b8b8;
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width:100%;
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padding: 50px;
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  clear:both;
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position: relative;
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bottom:0;
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border-top: solid 8px #8cff00;
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}
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footer p {
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margin:auto;
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padding-top:10px;
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text-align:center;
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width:50%;
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color:#006600;
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.firstHeading {
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display:none;
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}
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table{
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color:#888;
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font-size:16px;
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border: solid 1px #ccc;
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}
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h1 {
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font-size: 26px;
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color:#339900;
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}
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h2 {
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font-size: 22px;
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color:#339900;
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h3 {
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font-size: 18px;
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color:#339900;
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h4 {
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font-size: 16px;
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color:#339900;
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.listHorizontal{
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    display:inline-block;
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    margin-left: 0.3em;
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}
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.listHorizontal li{
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}
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div#globalWrapper {
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position: relative;
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font-size: 100%;
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width: 100%;
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margin: 0;
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padding: 0;
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padding-bottom: 10px;
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}
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</style>
   </head>
   </head>
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         <li><a href="https://2014.igem.org/Team:LIKA-CESAR-Brasil/Safety">SAFETY</a></li>
         <li><a href="https://2014.igem.org/Team:LIKA-CESAR-Brasil/Safety">SAFETY</a></li>
         <li><a href="https://2014.igem.org/Team:LIKA-CESAR-Brasil/Atribuitons">ATRIBUITIONS</a></li>
         <li><a href="https://2014.igem.org/Team:LIKA-CESAR-Brasil/Atribuitons">ATRIBUITIONS</a></li>
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        <li class="igem-logo"><a href="https://2014.igem.org/"></a></li>
             </ul>
             </ul>
           </div><!--/.nav-collapse -->
           </div><!--/.nav-collapse -->

Latest revision as of 02:13, 18 October 2014

LIKA | CESAR

NOTEBOOK

Transformation protocol

  • 1) Transform cells with 50 l of 1 µl plasmid.
  • 2) Dilute plasmid NEB buffer.
  • 3) Incubate the mixture of cells with the resistance plasmid for 30 minutes on ice.
  • 4) After 30 minutes, bring the sample to heat shock in a water bath at 42 ° C for 60 seconds, after adding 200 l SOC medium and incubate at 37 ° C for 2 hours. (Procedures performed in 2 ml tubes).
  • 5) Tagging cells transformed with plasmid resistance on solid LB containing antibiotics (chloramphenicol).
  • 6) 250 L of cell solution was plated on solid medium for 16 hours in an oven at 37 ° C microbiological.
  • 7) After the 16 hours incubation the counting antibiotic-resistant colonies was made (observed the efficiency of transformation).
  • 8) Calculation of transformation efficiency: Efficiency of transformation (dilution factor = 15) x colony count x 105 / μgDNA.
Marca LIKA Marca CESAR

Retrieved from "http://2014.igem.org/Team:LIKA-CESAR-Brasil/TransformationProtocol"