Team:LZU-China/wetlab2

From 2014.igem.org

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/*This is wetlab2*/
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<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" " http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
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<html xmlns=" http://www.w3.org/1999/xhtml">
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<head>
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<meta http-equiv="Content-Type" content="text/html; charset=gb2312" />
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<title>LZU-China 2014</title>
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<style type="text/css">
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body,ul,li,ol{list-style:none;padding:0px;margin:0px;font-size:12px;font-family:微软雅黑;}
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.picList .col img{width:100%;}
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.msg{float:left;width:900px;position:relative;left:50%;margin:30px 0 0 -450px;font-size:14px;}
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.msg{float:left;width:900px;position:relative;left:50%;margin:30px 0 0 -450px;font-size:14px;}
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.picList6 .col img{width:100%;}
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.picList6 .col a{display:block;width:580;height:377px;top:357px;background:rgba(0,0,0,0.7);color:white;font-style:normal;line-height:25px;padding:10px;opacity:0.8;text-decoration:none;}
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.msg{float:left;width:900px;position:relative;left:50%;margin:30px 0 0 -450px;font-size:14px;}
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.picList7{width:100%;float:left;}
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.picList7 .col{width:600px;height:397px;position:relative;overflow:hidden;float:left;margin:10px 0 0 10px;}
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.picList7 .col img,.col a{position:absolute;top:0px;left:0px;}
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.picList7 .col img{width:100%;}
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.picList7 .col a{display:block;width:580;height:377px;top:357px;background:rgba(0,0,0,0.7);color:white;font-style:normal;line-height:25px;padding:10px;opacity:0.8;text-decoration:none;}
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.msg{float:left;width:900px;position:relative;left:50%;margin:30px 0 0 -450px;font-size:14px;}
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.picList8 .col{width:600px;height:397px;position:relative;overflow:hidden;float:left;margin:10px 0 0 10px;}
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.picList8 .col img{width:100%;}
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.picList8 .col a{display:block;width:580;height:377px;top:357px;background:rgba(0,0,0,0.7);color:white;font-style:normal;line-height:25px;padding:10px;opacity:0.8;text-decoration:none;}
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.msg{float:left;width:900px;position:relative;left:50%;margin:30px 0 0 -450px;font-size:14px;}
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*{margin:0px;padding:0px;}
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body{background:#b1b1b1;margin:0px;padding:0px;font-size:14px;color:#000;}
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b#p1, b#p2, b#p3, b#p4, b#p5 {display:none;}
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#accordian {padding:0; margin:20px auto; list-style:none; width:730px; height:300px; overflow:hidden; position:relative; border:1px solid #682;}
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#accordian div {width:530px; height:300px; position:relative;}
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#accordian div span a {color:#fff;}
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padding-top: 100px;
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margin-bottom: 20px;
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font-family: Georgia, Times, "Times New Roman", serif;
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}
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h1, h2, h3, h4, h5 { font-family: Georgia, Times, "Times New Roman", serif;}
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#top-section { /*-- styling for default menu bar (edit, page, history, etc.) --*/
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background-color: #383838;
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border: 0 none;
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z-index: 100;
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top: 0;
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position: fixed;
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width: 975px;
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left: 50%;
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margin-left: -487px;
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}
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#top-section-bar { /*-- styling full width bar which hides behind default menu bar (edit, page, history, etc.) --*/
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background-color: #383838;
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height: 14px;
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display: block;
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z-index: 10;
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position: fixed;
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width: 100%;
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top: 0;
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}
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#menubar a:link, #menubar a:active, #menubar a:visited, #menubar a:hover, #menubar:hover { /*-- styling for default menu bar links (edit, page, history, etc.) --*/
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color: #727272;
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text-decoration: none;
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background-color: transparent;
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}
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body {
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background-color: #fff;
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}
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#globalWrapper, #content { /*-- changes default wiki settings --*/
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width: 100%;
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height: 100%;
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border: 0px;
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background-color: transparent;
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padding: 0px;
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}
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html, body, .wrapper { /*-- changes default wiki settings --*/
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width: 100%;
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height: 100%;
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background-color: transparent;
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}
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#contentcontainer { /*-- creates container for all content on page --*/
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font-family: Arial, Helvetica, sans-serif;
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font-weight: normal;
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font-size: 14px;
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color: #414141;
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width: 960px;
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margin-left: auto;
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margin-right: auto;
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background-color: #transparent;
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margin-top: 0px;
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}
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.sidemenu, .sidemenu li {
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list-style-type: none;
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list-style-image: none;
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font-family: verdana;
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text-decoration: none;
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color:#000;
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font-size: 14px;
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}
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.sidemenu li {display: block;}
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.sidemenu a {
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text-decoration:none;
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color: #000;
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width: 180px;
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height: 100%;
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padding: 3px 5px;
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transition: .25s ease-in-out;
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-moz-transition: .25s ease-in-out;
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-webkit-transition: .25s ease-in-out;
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}
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.sidemenu a.greyout { color: #909090; }
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.sidemenu a:hover {
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color:#666;
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background-color: #d9f5aa;
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}
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.sub-sidemenu {font-size: 10px; margin-bottom: 20px; color: #666; display: none;}
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.sub-sidemenu a {color: #666;}
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div.underconst
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{
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padding:10px;
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border-radius:5px;
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background-color:pink;
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text-align:center;
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}
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a.anchor{display: block; position: relative; top: -80px; visibility: hidden;}
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.contentpara {margin-bottom: 30px;}
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span.email{font-family: monospace; font-weight: normal;}
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fieldset {border: 1px solid #337f53;}
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.greyout {color: #A0A0A0;}
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.highlightme {background-color: #FFFF00;}
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#alertContainer {border: 2px solid red; margin-bottom: 10px; width: 936px;}
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#annContainer {margin-left: 8px;}
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#newsContainer {border: 1px solid #ccc;}
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.newsTitle {
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display: block;
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color: #414141;
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font-size: 25px; 
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        font-family: Georgia, Times, "Times New Roman", serif;
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padding: 10px 15px 5px 10px;
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border-bottom: 1px solid #ccc;
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margin-bottom: 0px;
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}
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.newsItem {
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border-bottom: 1px solid #ccc;
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display: block;
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padding: 5px 15px 0px 10px;
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margin-bottom: 0px;
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}
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.newsItem h3 {
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width: auto;
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display: inline;
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font-size: 14px;
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font-family: Arial, Helvetica, sans-serif;
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padding: 0px;
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margin: 0px;
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}
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.newsItem img {
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float: right;
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clear: right;
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width: 80px;
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padding-left: 15px;
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display: inline-block;
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}
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.newsItem .newsDate {
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font-style: italic;
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font-size: 14px;
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display: inline-block;
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color: #999;
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float: right;
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padding: 0px 0px 10px 15px;
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.newsItem p, .newsItem ul, .newsItem li {
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padding: 0px;
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margin-left: 20px;
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font-size: 12px;
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line-height: 1.2;
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.newsItem p {margin-top: 10px;}
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.annItem {
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border: 2px solid #49b677;
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display: block;
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padding: 0px 15px 10px 10px;
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margin-bottom: 18px;
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position: relative;
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height: 160px;
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vertical-align: middle;
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}
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.annItem .annCentered {
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display: table-cell;
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vertical-align: middle;
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height: 160px;
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}
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.annCentered h3 {
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padding: 0px;
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display: block;
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margin-left: auto;
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margin-right: auto;
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text-align: center;
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}
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.annItem ul, .annItem li {padding: 0px; margin: 0px;}
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.annItem li {list-style: none; margin: 5px}
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@charset "utf-8";
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body,h1,h2,h3,h4,h5,h6,hr,p,blockquote,dl,dt,dd,ul,ol,li,pre,form,fieldset,legend,button,input,textarea,th,td,figure{
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padding:0;
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font-size: 14px;
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text-align: left;
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}
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html{background-color:#e9ecec}
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table{table-layout:fixed;border-collapse:collapse}
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section,article,aside,header,footer,nav,dialog,figure{display:block}
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body,button,input,select,textarea,td,th{color:#333;font:12px/1.5 tahoma,arial,\5b8b\4f53}
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h1,h2,h3,h4,h5,h6{font-size:100%}
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small{font-size:12px}ul,ol{list-style:none}a{text-decoration:none;color:#06C}
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a:hover{text-decoration:underline;color:#06C}
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sup{vertical-align:text-top}
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sub{vertical-align:text-bottom}
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legend{color:#000}
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fieldset,img{border:0 none}
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button,input,select,textarea{font-size:100%}
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table{
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border-collapse:collapse;
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border-spacing:0;
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text-align: center;
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input:focus{outline:none}
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.hd-main{height:60px;background:#fff;z-index:15;border-bottom:#b3b3b3 1px solid;box-shadow:0 0 5px #333}
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.f-icon{background-image:url('http://www.codefans.net/jscss/demoimg/201403//frame-icon.png');background-repeat:no-repeat}
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.hd-main{padding:0 5px;height:49px;min-width:800px;background:#252525;border-bottom:0}
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.hd-main .has-pulldown{cursor:pointer;position:relative;*z-index:10000}
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.hd-main .pulldown{position:absolute;cursor:default;display:none;top:30px;left:0}
 +
.hd-main .pulldown .arrow{*margin-bottom:-1px;_margin-bottom:0;height:0;width:0;font-size:0;line-height:0;border-width:7px;border-style:solid;border-color:transparent transparent rgba(255, 255, 255,0.8) transparent;_filter:chroma(color=tomato);_border-color:tomato tomato rgba(255, 255, 255,0.8) tomato;position:relative;display:block;left:20px;z-index:7}
 +
.hd-main .pulldown .content{background:rgba(255, 255, 255,0.8);color:#333;text-align:left;border-radius:3px;border:rgb(175,175,175) 1px solid;border-width:0 1px 1px 1px;box-shadow:0 2px 4px rgba(0,0,0,.2);position:relative;z-index:6}
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.hd-main .pulldown-canvas{position:absolute;top:0;left:0;width:100%;height:100%;opacity:0;filter:alpha(opacity=0)}
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.hd-main .logo{background-image:url('https://static.igem.org/mediawiki/2014/7/72/2014logomin.png');background-repeat:no-repeat;display:block;height:36px;width:175px;float:left;margin-left:10px;background-position:0 0;margin-top:5px;display:inline}
 +
.hd-main .navs{padding:0 0 3px 25px;height:45px;float:left}
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.hd-main .navs a,.hd-main .info a,.hd-main .navs a:visited,.hd-main .info a:visited,.hd-main .navs a:active,.hd-main .info a:active{color:#d8d8d8}
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.hd-main .navs a:hover,.hd-main .navs a:active{text-decoration:none;background-color:#0f0f0f}
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.hd-main .navs .def-nav,.hd-main .navs .pulldown-nav,.hd-main .navs .separate{display:block;float:left;height:48px;font:18px/30px "Microsoft YaHei","Microsoft JhengHei";color:#d8d8d8;text-align:center;width:125px;line-height:48px}
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.hd-main .navs .def-nav .app-url{left:185px;width:425px;top:48px}
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.hd-main .navs .def-nav .pulldown-canvas-url{width:575px;height:109px}
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.hd-main .navs .def-nav .app-url .arrow{left:140px}
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.hd-main .navs .app-url .content{padding:17px 0 0 22px;height:91px}
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.hd-main .navs .app-url .content a,.hd-main .navs .app-url .content a:hover{color:#333}
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.hd-main .navs .app-url .li{cursor:pointer;padding-top:8px;width:66px;height:66px;border-radius:3px;display:block;float:left;text-align:center}
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.hd-main .navs .app-url .li:hover{background-color:#e0e1e5}
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.hd-main .navs .app-url .li .icon{margin:0 auto;width:50px;height:50px;display:block;background:url('https://static.igem.org/mediawiki/2014/9/94/Icons50X50nb.png') no-repeat}
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.hd-main .navs .app-url .li:hover .icon{background-image:url('https://static.igem.org/mediawiki/2014/9/94/Icons50X50nb.png')}
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.hd-main .navs .app-url .tongxunlu .icon{background-position:-50px 0}
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.hd-main .navs .app-url .xiangce .icon{background-position:-100px 0}
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      <td width="1201" height="265" rowspan="2"><p><img src="https://static.igem.org/mediawiki/2014/thumb/9/99/9.notebook.top.jpg/800px-9.notebook.top.jpg" width="1366" height="270" /></p></td>
 +
      <td width="100%" height="199" bgcolor="#187ADB">&nbsp;</td>
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      <td width="25%">&nbsp;</td>
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      <td><p>&nbsp;</p>
 +
        <p>&nbsp;</p>
 +
        <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&quot;Rome was not built in a day.&quot; No matter which group we belong to, we took our work for almost a whole year. What did we do on earth? This notebook will show you in detail.</p>
 +
        <p>&nbsp;</p>
 +
        <p>&nbsp;</p>
 +
        <div><a href="https://2014.igem.org/Team:LZU-China/Notebook"><img src="https://static.igem.org/mediawiki/2014/8/8f/Z.wabb%281%29.png" width="214" height="99" /></a> <a href="https://2014.igem.org/Team:LZU-China/NotebookB"><img src="https://static.igem.org/mediawiki/2014/e/e6/Z.wabb%282%29.png" width="215" height="99" /></a><a href="https://2014.igem.org/Team:LZU-China/NotebookC"><img src="https://static.igem.org/mediawiki/2014/0/0e/Z.wabb%283%29.png" width="212" height="99" /></a></div>
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      <p>&nbsp;</p></td>
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      <td width="25%">&nbsp;</td>
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<div id="apDiv6"><img src="https://static.igem.org/mediawiki/2014/6/6e/Lzulogo.png" width="220" height="221" /></div>
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  <table width="100%" border="0">
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      <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg"><p>&nbsp;</p>
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        <p>&nbsp;</p>
 +
        <table width="100%" border="0"  >
 +
        <tr>
 +
          <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg" >&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;THE NOTEBOOK OF WET LAB GROUP A</td>
 +
        </tr>
 +
      </table></td>
 +
    </tr>
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 +
      <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg" ><table width="100%" border="0">
 +
        <tr>
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          <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg"  width="30%">&nbsp;</td>
 +
          <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg"  width="57%"><p>&nbsp;</p>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2014.5.16-5.23<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The pre-experience of the growth of three  strains(<em>Shewanella onedensis,</em> <em>Pseudomonas aeruginosa </em>and LZU-3, which is separated in Li&rsquo;s  lab )in different PNP concentration and  Cr(VI) concentration.</p>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2014/f/f5/Z.da1.png" width="700" height="173" /></p>
 +
            <div><img src="file:///E|/[iGEM]/wiki/2014logomin.png" width="358" height="40" /></div>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="file:///E|/https://static.igem.org/mediawiki/2014/e/e0/Partpic2.png" width="301" height="251" /></p>
 +
            <div><img src="file:///E|/[iGEM]/wiki/partpic2.png" width="301" height="251" /></div>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2014.5.24-5.31<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Measurement for three strains&rsquo;(<em>Shewanella onedensis,</em> <em>Pseudomonas aeruginosa </em>and LZU-3, which is separated in Li&rsquo;s  lab ) Cr(VI) reduction activity.</p>
 +
            <div><img src="file:///E|/[iGEM]/wiki/dblbluebar.jpg" alt="" width="1365" height="463" /></div>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2014/thumb/d/dc/Z.a2.png/463px-Z.a2.png" width="467" height="605" /></p>
 +
<p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2014.6.1-2014.6.8<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;1.Another measurement for three strains&rsquo;(as above  said) Cr(VI) reduction activity.</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2014/a/a6/Z.a3.png" width="569" height="482" /></p>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2.Construction of the phylogeny tree containing the LZU-3 </p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2014/5/52/Z.4.png" width="654" height="503" /></p>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2014.6.8-2014.7.24<br />
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The PCR cycling for BBa_I723020 and BBa_I723133</p>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;1. The primers(synthesized by BGI,shenzhen)</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;1.1 Promoter of BBa_I723133 <br />
 +
              The forward primer (5&rsquo;  TATACTAGTCTGGGGCGAGAGGCGACGAC 3&rsquo;) contained a SpeI cutsite <br />
 +
              The reverse primer (5&rsquo;  GGGGAATTCTAGAATGTGGGCTGCTTGGTG 3&rsquo;) contained a EcoRI and XbaI cutsite. <br />
 +
              2313bp</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;1.2 BBa_I723020<br />
 +
              The forward primer  (5&rsquo;GGTGCTGCAGACTAGTATTGAAGGGTCACCACTATTTTTATTTTA 3&rsquo;) containes SpeI and PstI  cutsites <br />
 +
              the reverse primer (5&rsquo;  ATGAATTCTCTAGACCTGCTGGAGGGCGTGAAC 3&rsquo;) containes EcoRI and XbaI cutsites. <br />
 +
              340bp</p>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2.The  PCR system</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;0.2&mu;L ExTaq.<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;1&mu;L bacterial liquid<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;1&mu;L dNTP<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;1&mu;L forward primer<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;1&mu;L reverse primer<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2.5&mu;L 10x Buffer<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;18.5&mu;L ddH2O</p>
 +
<p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2014.7.31-8.1<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Pre-experience for the function of BBa_K381001.<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;BBa_K381001 transformation to E.coli DH5 alpha<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;There are two groups to test the green fluorescence under microscope:<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;(These groups' total sizes are all 20&mu;L, had been shaken for 2hours before observation,400x)<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;a.bacterial liquid with 10mM PNP concentration.<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;b.bacterial liquid with 20mM PNP concentration.<br />
 +
            </p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2014/2/2d/Z.5.png" width="704" height="307" /></p>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2014.8.2-2014.8.6<br />
 +
              Pre-experience for the function of BBa_K381001.<br />
 +
              BBa_K381001 transformation to E.coli DH5 alpha<br />
 +
              There are four groups to test the green fluorescence under microscope:<br />
 +
              (These groups' total sizes are all 20&mu;L, had been shaken for 2hours before observation,1000x)<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;a.bacterial liquid with 10mM PNP concentration.b.<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;b.bacterial liquid with ddH2O<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;c.bacterial liquid with 10mM KNO3<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;d.bacterial liquid with 10mM KCl.<br />
 +
            </p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2014/4/41/Z.a6.png" width="455" height="340" /></p>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2014.8.25-9.21<br />
 +
              &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;1.Construction of BBa_K1523101<br />
 +
            </p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2014/2/28/Z.a7.png" width="489" height="90" /></p>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The size of the whole part is about 3700bp(without plasmid), we test the assembling result by PCR(sense:5'---TTCCCATCTATAATCCTCCCTGATTCTTCG---3';anti-sense:5'---GAATTCTCTAGATTACAACTGTTGTTCAAGCTGTT---3'). From the gel picture we can see the size is right.</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2014/c/cf/Z.a8.png" width="643" height="305" /></p>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2.Construction of BBa_K1523024(yieF) and BBa_K1523005(chrR)</p>
 +
<p>&nbsp;</p>
 +
<p>&nbsp;</p>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2014.9.21-10.2<br />
 +
  &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Construction of BBa_K1523021(<em>nahR</em>),  K1523022(<em>nahE</em>) and K1523023(<em>nahG</em>).</p>
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Revision as of 03:01, 17 October 2014

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" " http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd"> LZU-China 2014

 
 
 

 

 

        "Rome was not built in a day." No matter which group we belong to, we took our work for almost a whole year. What did we do on earth? This notebook will show you in detail.

 

 

 

 

 

 

 
 
 

 

 

 

        THE NOTEBOOK OF WET LAB GROUP A
 

 

 

        2014.5.16-5.23
        The pre-experience of the growth of three strains(Shewanella onedensis, Pseudomonas aeruginosa and LZU-3, which is separated in Li’s lab )in different PNP concentration and Cr(VI) concentration.

 

        

 

                                        

 

 

        2014.5.24-5.31
        Measurement for three strains’(Shewanella onedensis, Pseudomonas aeruginosa and LZU-3, which is separated in Li’s lab ) Cr(VI) reduction activity.

 

                                

 

        2014.6.1-2014.6.8
        1.Another measurement for three strains’(as above said) Cr(VI) reduction activity.

                 

 

        2.Construction of the phylogeny tree containing the LZU-3

             

 

 

          2014.6.8-2014.7.24
          The PCR cycling for BBa_I723020 and BBa_I723133

 

       1. The primers(synthesized by BGI,shenzhen)

       1.1 Promoter of BBa_I723133
The forward primer (5’ TATACTAGTCTGGGGCGAGAGGCGACGAC 3’) contained a SpeI cutsite
The reverse primer (5’ GGGGAATTCTAGAATGTGGGCTGCTTGGTG 3’) contained a EcoRI and XbaI cutsite.
2313bp

       1.2 BBa_I723020
The forward primer (5’GGTGCTGCAGACTAGTATTGAAGGGTCACCACTATTTTTATTTTA 3’) containes SpeI and PstI cutsites
the reverse primer (5’ ATGAATTCTCTAGACCTGCTGGAGGGCGTGAAC 3’) containes EcoRI and XbaI cutsites.
340bp

 

      2.The PCR system

            0.2μL ExTaq.
            1μL bacterial liquid
            1μL dNTP
            1μL forward primer
            1μL reverse primer
            2.5μL 10x Buffer
            18.5μL ddH2O

 

          2014.7.31-8.1
      Pre-experience for the function of BBa_K381001.
      BBa_K381001 transformation to E.coli DH5 alpha
      There are two groups to test the green fluorescence under microscope:
      (These groups' total sizes are all 20μL, had been shaken for 2hours before observation,400x)
                  a.bacterial liquid with 10mM PNP concentration.
                  b.bacterial liquid with 20mM PNP concentration.

          

 

          2014.8.2-2014.8.6
Pre-experience for the function of BBa_K381001.
BBa_K381001 transformation to E.coli DH5 alpha
There are four groups to test the green fluorescence under microscope:
(These groups' total sizes are all 20μL, had been shaken for 2hours before observation,1000x)
      a.bacterial liquid with 10mM PNP concentration.b.
      b.bacterial liquid with ddH2O
      c.bacterial liquid with 10mM KNO3
      d.bacterial liquid with 10mM KCl.

                              

 

          2014.8.25-9.21
      1.Construction of BBa_K1523101

                              

 

      The size of the whole part is about 3700bp(without plasmid), we test the assembling result by PCR(sense:5'---TTCCCATCTATAATCCTCCCTGATTCTTCG---3';anti-sense:5'---GAATTCTCTAGATTACAACTGTTGTTCAAGCTGTT---3'). From the gel picture we can see the size is right.

             

 

      2.Construction of BBa_K1523024(yieF) and BBa_K1523005(chrR)

 

 

          2014.9.21-10.2
      Construction of BBa_K1523021(nahR), K1523022(nahE) and K1523023(nahG).