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|
| Line 2: |
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| | <head> | | <head> |
| | <meta http-equiv="Content-Type" content="text/html;charset=UTF-8" /> | | <meta http-equiv="Content-Type" content="text/html;charset=UTF-8" /> |
| - | <title>UESTC-China</title> | + | <title>Plant Vs HCHO</title> |
| | + | |
| | + | |
| | <style> | | <style> |
| | | | |
| Line 47: |
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| | #header{ | | #header{ |
| | height: 280px; | | height: 280px; |
| - | background-image:url("https://static.igem.org/mediawiki/2014/a/a3/Newbanner.jpg"); | + | background-image:url("https://static.igem.org/mediawiki/2014/a/a3/Newbanner.jpg");/* yan change to plant-vs HCHO*/ |
| | background-position: top center; | | background-position: top center; |
| | background-repeat: no-repeat; | | background-repeat: no-repeat; |
| Line 70: |
Line 72: |
| | .top-menu-each{ | | .top-menu-each{ |
| | height: 63px; | | height: 63px; |
| | + | |
| | font-family: helveticallt,"微软雅黑"; | | font-family: helveticallt,"微软雅黑"; |
| | color: white; | | color: white; |
| Line 75: |
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| | float: left; | | float: left; |
| | cursor: pointer; | | cursor: pointer; |
| - | font-weight: bold; | + | font-weight: bold; |
| | } | | } |
| | | | |
| Line 86: |
Line 89: |
| | .arrow{ | | .arrow{ |
| | float: left; | | float: left; |
| - | background-image: url("http://fudanigem.phx-2.ifreeurl.com/img/arrow.png"); | + | background-image: url("http://fudanigem.phx-2.ifreeurl.com/img/arrow.png");/*needn't change*/ |
| | line-height: 63px; | | line-height: 63px; |
| | width: 12px; | | width: 12px; |
| Line 150: |
Line 153: |
| | padding-left: 35px; | | padding-left: 35px; |
| | padding-right: 10px; | | padding-right: 10px; |
| - | background-image: url('http://fudanigem.phx-2.ifreeurl.com/img/big-photo-hover.png'); | + | background-image: url('http://fudanigem.phx-2.ifreeurl.com/img/big-photo-hover.png');/*no change*/ |
| | background-repeat: no-repeat; | | background-repeat: no-repeat; |
| | margin-top: -130px; | | margin-top: -130px; |
| Line 168: |
Line 171: |
| | font-size: 30px; | | font-size: 30px; |
| | line-height: 30px; | | line-height: 30px; |
| - | font-family: helveticallt,"微软雅黑"; | + | font-family: helveticalmd,"微软雅黑"; |
| | font-weight: bold; | | font-weight: bold; |
| | padding-top: 25px; | | padding-top: 25px; |
| Line 210: |
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| | | | |
| | height: 180px; | | height: 180px; |
| - | background-color: #394264; | + | background-color:white; |
| | border-top-left-radius:8px; | | border-top-left-radius:8px; |
| | border-top-right-radius:8px; | | border-top-right-radius:8px; |
| Line 229: |
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| | height: 60px; | | height: 60px; |
| | background-color: #394264; | | background-color: #394264; |
| - | font-family: helveticallt,"微软雅黑"; | + | font-family: ubuntu,"宋体"; |
| | font-size: 18px; | | font-size: 18px; |
| | line-height: 60px; | | line-height: 60px; |
| Line 254: |
Line 257: |
| | #middle-photo{ | | #middle-photo{ |
| | float: left; | | float: left; |
| - | margin-bottom: 25px; | + | margin-bottom: 445px; |
| | } | | } |
| | .middle-photo-each{ | | .middle-photo-each{ |
| - | padding:0px 0px 0px 0px; | + | padding:9px 9px 9px 9px; |
| | background-color: white; | | background-color: white; |
| | margin-bottom: 15px; | | margin-bottom: 15px; |
| - | margin-left: 0px; | + | margin-left: 250px; |
| | | | |
| | } | | } |
| Line 277: |
Line 280: |
| | color: black; | | color: black; |
| | font-size: 16px; | | font-size: 16px; |
| - | font-family: helveticallt,"微软雅黑"; | + | font-family: "微软雅黑"; |
| | line-height: 20px; | | line-height: 20px; |
| | } | | } |
| Line 291: |
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| | } | | } |
| | #top{ | | #top{ |
| - | background-image:url('http://fudanigem.phx-2.ifreeurl.com/img/top.png'); | + | background-image:url('http://fudanigem.phx-2.ifreeurl.com/img/top.png');/*no change*/ |
| | width: 57px; | | width: 57px; |
| | height: 57px; | | height: 57px; |
| Line 301: |
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| | #logo{ | | #logo{ |
| | position: absolute; | | position: absolute; |
| - | background-image: url("https://static.igem.org/mediawiki/2014/9/99/NewLogo.png"); | + | background-image: url("https://static.igem.org/mediawiki/2014/9/99/NewLogo.png");/*no change*/ |
| | width: 154px; | | width: 154px; |
| | height: 147px; | | height: 147px; |
| Line 321: |
Line 324: |
| | color:white; | | color:white; |
| | } | | } |
| | + | #intro1{ |
| | + | font-family: helveticalmd,"微软雅黑"; |
| | + | font-size: 50px; |
| | + | margin-top: 40px; |
| | + | margin-left: 10px; |
| | + | color: white; |
| | + | } |
| | + | #intro2{ |
| | + | font-family: helveticalmd,"微软雅黑"; |
| | + | font-size: 20px; |
| | + | margin-left: 10px; |
| | + | color:white; |
| | + | } |
| | + | #introduction{ |
| | + | background-color:#1bbc9b |
| | + | } |
| | + | /*rocket-to-top*/ |
| | + | *{list-style:none;border:0;} |
| | | | |
| | + | #rocket-to-top div { |
| | + | left: 0; |
| | + | margin: 0; |
| | + | overflow: hidden; |
| | + | padding: 0; |
| | + | position: absolute; |
| | + | top: 0; |
| | + | width: 149px; |
| | + | } |
| | + | #rocket-to-top .level-2 { |
| | + | background: url("https://static.igem.org/mediawiki/2014/a/a6/UestcUp.png") no-repeat scroll -149px 0 transparent; |
| | + | display: none; |
| | + | height: 250px; |
| | + | opacity: 0; |
| | + | z-index: 1; |
| | + | } |
| | + | #rocket-to-top .level-3 { |
| | + | background: none repeat scroll 0 0 transparent; |
| | + | cursor: pointer; |
| | + | display: block; |
| | + | height: 150px; |
| | + | z-index: 2; |
| | + | } |
| | + | #rocket-to-top { |
| | + | background: url("https://static.igem.org/mediawiki/2014/a/a6/UestcUp.png") no-repeat scroll 0 0 transparent; |
| | + | cursor: default; |
| | + | display: block; |
| | + | height: 250px; |
| | + | margin: -125px 0 0; |
| | + | overflow: hidden; |
| | + | padding: 0; |
| | + | position: fixed; |
| | + | right: 0; |
| | + | top: 80%; |
| | + | width: 149px; |
| | + | z-index: 11; |
| | + | } |
| | + | /*rocket-to-top end*/ |
| | | | |
| | + | </style> |
| | + | </head> |
| | + | <body> |
| | + | <div id="total-wrap"> |
| | + | <div id="header"> |
| | + | <div id="emp"> |
| | + | <div id='logo'><a href="https://igem.org/Main_Page"><img src="https://static.igem.org/mediawiki/2014/6/60/LiLogo.png"></a></div> |
| | + | </div> |
| | + | <div id="top-menu"> |
| | + | <div class="top-menu-each"> |
| | + | <a href="https://2014.igem.org/Team:UESTC-China"><p>Home</p></a> |
| | + | </div> |
| | + | <div class="top-menu-each"> |
| | + | <div class="arrow"></div> |
| | + | <p>Team</p> |
| | + | <div class="top-menu-sub"> |
| | + | <ul> |
| | | | |
| - | #SensorEditingArea{position:absolute; left:0px; top:300px; height:6500px; width:1213px; background-color:#ffffff; padding:60px 0px 0px 0px;}
| |
| | | | |
| | + | <a href="https://2014.igem.org/Team:UESTC-China/Team"><li>Members</li></a> |
| | + | <a href="https://2014.igem.org/Team:UESTC-China/Attribution"><li>Attribution</li></a> |
| | + | <a href="https://igem.org/Team.cgi?year=2014&team_name=UESTC-China"><li>Team Profile</li></a> |
| | + | <a href="https://2014.igem.org/Team:UESTC-China/Notebook"><li>Notebook</li></a> |
| | + | </ul> |
| | + | </div> |
| | + | </div> |
| | | | |
| - | .SensorEditingAreaClass p{position:relative;left:0px; width:1140px; font-size:23px; font-family: calibri, arial, helvetica, sans-serif; }
| |
| - | .SensorEditingAreaClass a{color:#1f8a70;}
| |
| | | | |
| - | #SectionTitles1{
| |
| - | position:relative;
| |
| - | left:30px;
| |
| - | background-color:#1f8a70;
| |
| - | padding:0 10px;
| |
| - | height:40px;
| |
| - | line-height:40px;
| |
| - | font-size:24px; font-family: calibri, arial, helvetica, sans-serif; border-bottom:0px; color:#ffffff; font-weight: bold;font-style: Italic; text-align:center; width:1140px; }
| |
| | | | |
| - | #SectionTitles2{
| + | <div class="top-menu-each"> |
| - | position:relative;
| + | <div class="arrow"></div> |
| - | top:350px; | + | <p>Project</p> |
| - | left:30px;
| + | <div class="top-menu-sub"> |
| - | background-color:#1f8a70;
| + | <ul> |
| - | padding:0 10px;
| + | <a href="https://2014.igem.org/Team:UESTC-China/Project"><li>Overview</li></a> |
| - | height:40px;
| + | <a href="https://2014.igem.org/Team:UESTC-China/Design"><li>Design</li></a> |
| - | line-height:40px;
| + | <a href="https://2014.igem.org/Team:UESTC-China/result"><li>Result</li></a> |
| - | font-size:24px; font-family: calibri, arial, helvetica, sans-serif; border-bottom:0px; color:#ffffff; font-weight: bold;font-style: Italic; text-align:center; width:1140px; }
| + | <a href="https://2014.igem.org/Team:UESTC-China/Futurework"> |
| | + | <li>Future work</li></a> |
| | | | |
| - | #SectionTitles3{
| + | </ul> |
| - | position:relative;
| + | </div> |
| - | left:30px;
| + | </div> |
| - | top: 720px; | + | <div class="top-menu-each"> |
| - | background-color:#1f8a70;
| + | <div class="arrow"></div> |
| - | padding:0 10px;
| + | <p>Modeling</p> |
| - | height:40px;
| + | <div class="top-menu-sub"> |
| - | line-height:40px;
| + | <ul> |
| - | font-size:24px; font-family: calibri, arial, helvetica, sans-serif; border-bottom:0px; color:#ffffff; font-weight: bold;font-style: Italic; text-align:center; width:1140px; }
| + | <a href="https://2014.igem.org/Team:UESTC-China/Modeling"><li>Modeling</li></a> |
| | + | </ul> |
| | + | </div> |
| | + | </div> |
| | + | <div class="top-menu-each"> |
| | + | <div class="arrow"></div> |
| | + | <p>Parts</p> |
| | + | <div class="top-menu-sub"> |
| | + | <ul> |
| | + | <a href="https://2014.igem.org/Team:UESTC-China/Material"><li>BioBrick</li></a> |
| | + | <a href="https://2014.igem.org/Team:UESTC-China/Protocol"><li>Protocol</li></a> |
| | | | |
| - | #SectionTitles4{
| + | </ul> |
| - | position:relative;
| + | </div> |
| - | top: 1100px; | + | </div> |
| - | left:30px;
| + | <div class="top-menu-each"> |
| - | background-color:#1f8a70;
| + | <div class="arrow"></div> |
| - | padding:0 10px;
| + | <p>Human Practice</p> |
| - | height:40px;
| + | <div class="top-menu-sub"> |
| - | line-height:40px;
| + | <ul> |
| - | font-size:24px; font-family: calibri, arial, helvetica, sans-serif; border-bottom:0px; color:#ffffff; font-weight: bold;font-style: Italic; text-align:center; width:1140px; }
| + | <a href="https://2014.igem.org/Team:UESTC-China/HumanPractice.html"><li>Knowledge quiz</li></a> |
| | + | <a href="https://2014.igem.org/Team:UESTC-China/Lecture.html"><li>Lecture</li></a> |
| | + | <a href="https://2014.igem.org/Team:UESTC-China/Communication.html"><li>Communication</li></a> |
| | + | <a href="https://2014.igem.org/Team:UESTC-China/Questionnaire.html"><li>Questionnaire</li></a> |
| | + | </ul> |
| | + | </div> |
| | + | </div> |
| | | | |
| - | #SectionTitles5{
| + | <div class="top-menu-each"> |
| - | position:relative;
| + | <div class="arrow"></div> |
| - | top: 1500px; | + | <p>Safety</p> |
| - | left:30px;
| + | <div class="top-menu-sub"> |
| - | padding:0 10px;
| + | <ul> |
| - | height:40px;
| + | <a href="https://2014.igem.org/Team:UESTC-China/Safety"><li>Safety</li></a> |
| - | line-height:40px;
| + | </ul> |
| - | font-size:24px; font-family: calibri, arial, helvetica, sans-serif; border-bottom:0px; color:#ffffff; font-weight: bold;font-style: Italic; text-align:center; width:1140px; }
| + | </div> |
| | + | </div> |
| | | | |
| - | #SectionTitles6{
| |
| - | position:relative;
| |
| - | top: 1850px;
| |
| - | left:30px;
| |
| - | background-color:#1f8a70;
| |
| - | padding:0 10px;
| |
| - | height:40px;
| |
| - | line-height:40px;
| |
| - | font-size:24px; font-family: calibri, arial, helvetica, sans-serif; border-bottom:0px; color:#ffffff; font-weight: bold;font-style: Italic; text-align:center; width:1140px; }
| |
| | | | |
| - | #SectionTitles7{
| |
| - | top: 2290px;
| |
| - | position:relative;
| |
| - | left:30px;
| |
| - | padding:0 10px;
| |
| - | height:40px;
| |
| - | line-height:40px;
| |
| - | font-size:24px; font-family: calibri, arial, helvetica, sans-serif; border-bottom:0px; color:#ffffff; font-weight: bold;font-style: Italic; text-align:center; width:1140px; }
| |
| | | | |
| - | #SectionTitles8{
| + | </div> |
| - | top: 2650px;
| + | </div> |
| - | position:relative;
| + | <div id="big-photo"> |
| - | left:30px;
| + | <div id="big-photo-wrap"> |
| - | background-color:#1f8a70;
| + | <img src="https://static.igem.org/mediawiki/2014/2/21/All_students1.jpg" alt="pic"> |
| - | padding:0 10px;
| + | <div id="big-photo-hover"> |
| - | height:40px;
| + | <p class="header">Our Team</p> |
| - | line-height:40px;
| + | <p class="content">Brilliant guys.</p> |
| - | font-size:24px; font-family: calibri, arial, helvetica, sans-serif; border-bottom:0px; color:#ffffff; font-weight: bold;font-style: Italic; text-align:center; width:1140px; }
| + | </div> |
| | + | </div> |
| | + | </div> |
| | + | <div id="introduction"> |
| | + | <p id="intro1"><b>Introduction</b></p> |
| | + | <br/> |
| | + | <p id="intro2">As the rapid economic development in China, the residential conditions of people have been greatly improved. Unfortunately, the potential harm of their indoor decoration was always neglected until people get sick. One of the most harmful gases of the indoor pollution could be the formaldehyde.<br/>Formaldehyde is a toxic gas appears in every single room which is newly decorated. It not only has bad smell, but also cause various of desease, like skin and mucous membrane irritation, immunity or memory decline, drowsiness, fatigue, cancer, allergic dermatitis.<br/>To decrease the concentration of HCHO, we got air purifier, physical adsorption, negative air ions, photocatalyst pathway and etc. We thought that the biological methods is the best. It’s delicate, it’s simple, and it’s safe. Therefore, we decided to cultivate a plant which got the ability to metabolize the formaldehyde. Actually, all kinds of plants can degrade the formaldehyde, but we decide to make their formaldehyde degradation system works better.<br/>In the future,we sincerely chrish the hope that our work will be put to use in our everyday life.</p> |
| | + | <br/> |
| | + | <img width="1215px" src="https://static.igem.org/mediawiki/2010/9/9c/Theory_Home-foot_UNR.png" alt="pic"> |
| | + | </div> |
| | + | |
| | + | <div id="footer"> |
| | + | |
| | + | <img width="1215px" height="180px" src='https://static.igem.org/mediawiki/2014/3/38/Uestcsponser.png' /> |
| | + | |
| | | | |
| - | #SectionTitles9{
| |
| - | top: 3000px;
| |
| - | position:relative;
| |
| - | left:30px;
| |
| - | padding:0 10px;
| |
| - | height:40px;
| |
| - | line-height:40px;
| |
| - | font-size:24px; font-family: calibri, arial, helvetica, sans-serif; border-bottom:0px; color:#ffffff; font-weight: bold;font-style: Italic; text-align:center; width:1140px; }
| |
| - | .textEditingArea{top:3400px;
| |
| - | position:absolute;
| |
| - | }
| |
| - | .textEditingTitle{
| |
| - | position:relative;
| |
| - | margin-right:20px;
| |
| - | background-color:#1f8a70;
| |
| - | left:30px;
| |
| - | padding:0 10px;
| |
| - | height:40px;
| |
| - | line-height:40px;
| |
| - | font-size:24px; font-family: calibri, arial, helvetica, sans-serif; border-bottom:0px;
| |
| - | color: #ffffff;text-align:center; width:1100px;
| |
| - | }
| |
| - | .textEditingstyle{
| |
| - | position:fixed;
| |
| - | color: #000000;line-height:40px;
| |
| - | font-size:24px; font-family:Microsoft Yahei; border-bottom:0px;
| |
| - | text-align:left; width:1140px; }
| |
| - |
| |
| - | #BiosensorsTable{position:absolute;margin-top: 20px; left:0px; width:1000px; font-family:calibri;arial;helvetica;sans-serif; font-size:18px; text-align:center; }
| |
| - | .mid{background: #FFFFCC;color: #797268; }
| |
| - | .top{background: #FFFF00; color: #797268; }
| |
| - | </style>
| |
| - | </head>
| |
| - | <body>
| |
| - | <div id="total-wrap">
| |
| - | <div id="header">
| |
| - | <div id="emp">
| |
| - | <div id='logo'><a href="https://igem.org"></a></div>
| |
| - | </div>
| |
| - | <div id="top-menu">
| |
| - | <div class="top-menu-each"> <a href="https://2014.igem.org/Team:UESTC-China">
| |
| - | <p>Home</p>
| |
| - | </a> </div>
| |
| - | <div class="top-menu-each">
| |
| - | <div class="arrow"></div>
| |
| - | <p>Team</p>
| |
| - | <div class="top-menu-sub">
| |
| - | <ul>
| |
| - | <a href="https://2014.igem.org/Team:UESTC-China/Team">
| |
| - | <li>Members</li>
| |
| - | </a> <a href="https://2014.igem.org/Team:UESTC-China/Attribution">
| |
| - | <li>Attribution</li>
| |
| - | </a> <a href="https://igem.org/Team.cgi?year=2014&team_name=UESTC-China">
| |
| - | <li>Team Profile</li>
| |
| - | </a> <a href="https://2014.igem.org/Team:UESTC-China/Notebook">
| |
| - | <li>Notebook</li>
| |
| - | </a>
| |
| - | </ul>
| |
| - | </div>
| |
| - | </div>
| |
| - | <div class="top-menu-each">
| |
| - | <div class="arrow"></div>
| |
| - | <p>Project</p>
| |
| - | <div class="top-menu-sub">
| |
| - | <ul>
| |
| - | <a href="https://2014.igem.org/Team:UESTC-China/Project">
| |
| - | <li>Overview</li>
| |
| - | </a> <a href="https://2014.igem.org/Team:UESTC-China/Design">
| |
| - | <li>Design</li>
| |
| - | </a> <a href="https://2014.igem.org/Team:UESTC-China/result">
| |
| - | <li>Result</li>
| |
| - | </a> <a href="https://2014.igem.org/Team:UESTC-China/Futurework">
| |
| - | <li>Future work</li>
| |
| - | </a>
| |
| - | </ul>
| |
| - | </div>
| |
| - | </div>
| |
| - | <div class="top-menu-each">
| |
| - | <div class="arrow"></div>
| |
| - | <p>Modeling</p>
| |
| - | <div class="top-menu-sub">
| |
| - | <ul>
| |
| - | <a href="https://2014.igem.org/Team:UESTC-China/Modeling">
| |
| - | <li>Modeling</li>
| |
| - | </a>
| |
| - | </ul>
| |
| - | </div>
| |
| - | </div>
| |
| - | <div class="top-menu-each">
| |
| - | <div class="arrow"></div>
| |
| - | <p>Parts</p>
| |
| - | <div class="top-menu-sub">
| |
| - | <ul>
| |
| - | <a href="https://2014.igem.org/Team:UESTC-China/Material">
| |
| - | <li>BioBrick</li>
| |
| - | </a> <a href="https://2014.igem.org/Team:UESTC-China/Protocol">
| |
| - | <li>Protocol</li>
| |
| - | </a>
| |
| - | </ul>
| |
| - | </div>
| |
| - | </div>
| |
| - | <div class="top-menu-each">
| |
| - | <div class="arrow"></div>
| |
| - | <p>Human Practice</p>
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| - | <div class="top-menu-sub">
| |
| - | <ul>
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| - | <a href="https://2014.igem.org/Team:UESTC-China/HumanPractice.html">
| |
| - | <li>Knowledge quiz</li>
| |
| - | </a> <a href="https://2014.igem.org/Team:UESTC-China/Lecture.html">
| |
| - | <li>Lecture</li>
| |
| - | </a> <a href="https://2014.igem.org/Team:UESTC-China/Communication.html">
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| - | <li>Communication</li>
| |
| - | </a> <a href="https://2014.igem.org/Team:UESTC-China/Questionnaire.html">
| |
| - | <li>Questionnaire</li>
| |
| - | </a>
| |
| - | </ul>
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| - | </div>
| |
| - | </div>
| |
| - | <div class="top-menu-each">
| |
| - | <div class="arrow"></div>
| |
| - | <p>Safety</p>
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| - | <div class="top-menu-sub">
| |
| - | <ul>
| |
| - | <a href="https://2014.igem.org/Team:UESTC-China/Safety">
| |
| - | <li>Safety</li>
| |
| - | </a>
| |
| - | </ul>
| |
| - | </div>
| |
| - | </div>
| |
| - | </div>
| |
| | </div> | | </div> |
| - | <div id="middle-photo">
| |
| - | <div id="SensorEditingArea" class="SensorEditingAreaClass">
| |
| - | <div class="tableEditing">
| |
| - | <div id="SectionTitles1" style="width:500px;">DNA ligation with T4 DNA Ligase
| |
| - | </h1>
| |
| - | <table border="1" id="BiosensorsTable" cellspacing="0";>
| |
| - | <tr>
| |
| - | <th class="top">Component</th>
| |
| - | <th class="top">20ul Reaction</th>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">1.33X Master Mix</td>
| |
| - | <td class="mid"> 15ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Insert DNA </td>
| |
| - | <td class="mid"> the moles of insert DNA to vector DNA is 5：1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">>Vector DNA</td>
| |
| - | <td class="mid"> 50ng</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">ddH2O</td>
| |
| - | <td class="mid"> To 20ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Incubate at 50℃ for 1 hour</td>
| |
| - | <td class="mid"></td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | </div>
| |
| - | <div id="SectionTitles2" style="width:500px;">DNA ligation with Gibson Assemble
| |
| - | </h1>
| |
| - | <table border="1" id="BiosensorsTable" cellspacing="0";>
| |
| - | <tr>
| |
| - | <th class="top">Component</th>
| |
| - | <th class="top">20ul Reaction </th>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Vector DNA</td>
| |
| - | <td class="mid">50ng</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Insert DNA</td>
| |
| - | <td class="mid"> the moles of insert DNA to vector DNA is 5：1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">10X T4 DNA ligase buffer</td>
| |
| - | <td class="mid">2ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">T4 DNA ligase</td>
| |
| - | <td class="mid">1ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">ddH2O</td>
| |
| - | <td class="mid">to 20ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Incubate at 25℃ for 12 hours</td>
| |
| - | <td class="mid"></td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | </div>
| |
| - | <div id="SectionTitles3" style="width:500px;">Double Digestion of DNA
| |
| - | </h1>
| |
| - | <table border="1" id="BiosensorsTable" cellspacing="0";>
| |
| - | <tr>
| |
| - | <th class="top">Component</th>
| |
| - | <th class="top">50ul Reaction </th>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">10x FastDigest buffer</td>
| |
| - | <td class="mid">5ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Restriction enzyme 1</td>
| |
| - | <td class="mid"> 1ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Restriction enzyme 2</td>
| |
| - | <td class="mid">1ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">DNA</td>
| |
| - | <td class="mid">1-2ug</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">ddH2O</td>
| |
| - | <td class="mid">to 50ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Incubate at 37℃ for 2 hours</td>
| |
| - | <td class="mid"></td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | </div>
| |
| - | <div id="SectionTitles4" style="width:500px;">PCR protocol for KOD-Plus-Neo
| |
| - | </h1>
| |
| - | <table border="1" id="BiosensorsTable" cellspacing="0";>
| |
| - | <tr>
| |
| - | <th class="top">Component</th>
| |
| - | <th class="top">50ul Reaction </th>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">ddH2O</td>
| |
| - | <td class="mid">33ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">10X buffer for KOD-Plus-Neo</td>
| |
| - | <td class="mid"> 5ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">MgSO4</td>
| |
| - | <td class="mid">1ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">10mM dNTPs</td>
| |
| - | <td class="mid">1ug</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">10uM forward primer</td>
| |
| - | <td class="mid">1ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">10uM reverse primer</td>
| |
| - | <td class="mid"> 1ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Template</td>
| |
| - | <td class="mid">1ul </td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">KOD enzyme</td>
| |
| - | <td class="mid"> 1ul</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | </div>
| |
| - | <div id="SectionTitles5" style="width:500px;">
| |
| - | <table border="1" id="BiosensorsTable" cellspacing="0";>
| |
| - | <tr>
| |
| - | <th class="top"></th>
| |
| - | <th class="top">Temperature </th>
| |
| - | <th class="top">Time </th>
| |
| - | <th class="top"> Cycle</th>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Step1</td>
| |
| - | <td class="mid">94℃</td>
| |
| - | <td class="mid">30s</td>
| |
| - | <td class="mid">X35 cycles</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid"></td>
| |
| - | <td class="mid"> 56℃</td>
| |
| - | <td class="mid">30s</td>
| |
| - | <td class="mid"></td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid"></td>
| |
| - | <td class="mid">68℃</td>
| |
| - | <td class="mid">30s/kb</td>
| |
| - | <td class="mid"></td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Step3</td>
| |
| - | <td class="mid">68℃</td>
| |
| - | <td class="mid">5min</td>
| |
| - | <td class="mid">X1 cycles</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid"></td>
| |
| - | <td class="mid">10℃</td>
| |
| - | <td class="mid">10min</td>
| |
| - | <td class="mid"></td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">10uM reverse primer</td>
| |
| - | <td class="mid"> 1ul</td>
| |
| - | <td class="mid">ddH2O</td>
| |
| - | <td class="mid">33ul</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | </div>
| |
| - | <div id="SectionTitles6" style="width:500px;">PCR protocol for KOD-Plus-Neo
| |
| - | </h1>
| |
| - | <table border="1" id="BiosensorsTable" cellspacing="0";>
| |
| - | <tr>
| |
| - | <th class="top">Component</th>
| |
| - | <th class="top">50ul Reaction </th>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">ddH2O</td>
| |
| - | <td class="mid">32ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">10X buffer for KOD-Plus-Neo</td>
| |
| - | <td class="mid"> 5ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">MgSO4</td>
| |
| - | <td class="mid">3ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">10mM dNTPs</td>
| |
| - | <td class="mid">5ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">10uM forward primer</td>
| |
| - | <td class="mid">1ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">10uM reverse primer</td>
| |
| - | <td class="mid"> 1ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Template</td>
| |
| - | <td class="mid">1ul </td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">KOD enzyme</td>
| |
| - | <td class="mid"> 1ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Each template will be diluted by the number of moles into 2-10ng/ul.</td>
| |
| - | <td class="mid"></td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | </div>
| |
| - | <div id="SectionTitles7" style="width:500px;">
| |
| - | <table border="1" id="BiosensorsTable" cellspacing="0";>
| |
| - | <tr>
| |
| - | <th class="top"></th>
| |
| - | <th class="top">Temperature </th>
| |
| - | <th class="top">Time </th>
| |
| - | <th class="top"> Cycle</th>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Step1</td>
| |
| - | <td class="mid">94℃</td>
| |
| - | <td class="mid">5min</td>
| |
| - | <td class="mid">X1 cycle</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Step2</td>
| |
| - | <td class="mid"> 94℃</td>
| |
| - | <td class="mid">30s</td>
| |
| - | <td class="mid">X35 cycles</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid"></td>
| |
| - | <td class="mid">56℃</td>
| |
| - | <td class="mid">30s</td>
| |
| - | <td class="mid"></td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid"></td>
| |
| - | <td class="mid">68℃</td>
| |
| - | <td class="mid">30s/kb</td>
| |
| - | <td class="mid"></td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Step3</td>
| |
| - | <td class="mid"> 68℃</td>
| |
| - | <td class="mid">5min</td>
| |
| - | <td class="mid">X1 cycles</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid"></td>
| |
| - | <td class="mid"> 10℃</td>
| |
| - | <td class="mid">10min</td>
| |
| - | <td class="mid"></td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | </div>
| |
| - | <div id="SectionTitles8" style="width:500px;">Colony PCR with Taq DNA polymerase
| |
| - | </h1>
| |
| - | <table border="1" id="BiosensorsTable" cellspacing="0";>
| |
| - | <tr>
| |
| - | <th class="top">Component</th>
| |
| - | <th class="top">25ul Reaction </th>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">ddH2O</td>
| |
| - | <td class="mid">15.8ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">10X Taq buffer</td>
| |
| - | <td class="mid"> 2.5ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">2.mM dNTPs</td>
| |
| - | <td class="mid">0.5ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">10uM forward primer</td>
| |
| - | <td class="mid">0.5ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">10uM reverse primer</td>
| |
| - | <td class="mid">0.5ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Taq enzyme</td>
| |
| - | <td class="mid"> 0.2ul</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Water with colony </td>
| |
| - | <td class="mid">5ul </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | </div>
| |
| - | <div id="SectionTitles9" style="width:500px;">
| |
| - | <table border="1" id="BiosensorsTable" cellspacing="0";>
| |
| - | <tr>
| |
| - | <th class="top"></th>
| |
| - | <th class="top">Temperature </th>
| |
| - | <th class="top">Time </th>
| |
| - | <th class="top"> Cycle</th>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Step1</td>
| |
| - | <td class="mid">95℃</td>
| |
| - | <td class="mid">5min</td>
| |
| - | <td class="mid">X1 cycles</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Step2</td>
| |
| - | <td class="mid"> 94℃</td>
| |
| - | <td class="mid">30s</td>
| |
| - | <td class="mid">X35 cycles</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid"></td>
| |
| - | <td class="mid">56℃</td>
| |
| - | <td class="mid">30s</td>
| |
| - | <td class="mid"></td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid"></td>
| |
| - | <td class="mid">72℃</td>
| |
| - | <td class="mid">1min/kb</td>
| |
| - | <td class="mid"></td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid">Step3</td>
| |
| - | <td class="mid">72℃</td>
| |
| - | <td class="mid">10min</td>
| |
| - | <td class="mid">X1 cycles</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td class="mid"></td>
| |
| - | <td class="mid">10℃</td>
| |
| - | <td class="mid">10min</td>
| |
| - | <td class="mid"></td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | </div>
| |
| - | </div>
| |
| - | <div class="textEditingArea" ><div class="textEditingTitle" style="width: 800px">E. coli Calcium Chloride competent cell protocol</br>
| |
| - | <p class="textEditingstyle">1）Streak E.coli cells (DH5a) on an LB plate; (BL21(DE3)LysS cells on LB plate+34 mg/ml chloramphenicol)<br>
| |
| - | 2） Allow cells to grow at 37℃ overnight<br>
| |
| - | 3）Place one colony in 10 ml LB media (+antibiotic selection if necessary), grow overnight at 37℃<br>
| |
| - | 4） Take 2 ml LB media and save for blank. Transfer 5 ml overnight DH5a culture into 500 ml LB media in 3 L flask<br>
| |
| - | 5） Allow cell to grow at 37℃ (250 rpm), until OD600= 0.4 (~2-3 hours)<br>
| |
| - | 6） Transfer cells to 2 centrifuge bottles (250 ml), and place cells on ice for 20 min<br>
| |
| - | 7） Centrifuge cells in at 4oC for 10 min at 3,000 g and subsequent resuspension may be done in the same bottle. Cells must remain cold for the rest of the procedure: Transport tubes on ice and resuspend on ice in the cold room<br>
| |
| - | 8） Pour off media and resuspend cells in 30 ml of cold 0.1 M CaCl2. Transfer the suspended cells into 50 ml polypropylene tubes, and incubate on ice for 30 min<br>
| |
| - | 9） Centrifuge cells at 4O℃ for 10 min at 3,000 g<br>
| |
| - | 10） Pour supernatant and resuspend cells (by pipetting) in 8 ml cold 0.1M CaCl2 containing 15% glycerol. Transfer 140 ml into (1.5 ml) Ependorff tubes placed on ice. Freeze the cells in liquid nitrogen. Cells stored at -80oC can be used for transformation for up to ~6 months<br>
| |
| - | 11） Add 10 to 40 ng (10 to 25 ml volume) of DNA to 250 ml of competent cells in step<br>
| |
| - | 12） Incubate the mixture on ice for 30 minutes.<br>
| |
| - | 13） Transfer the reaction to a 42℃ water for 1min.<br>
| |
| - | 14） Add 0.9 ml of LB culture to each tube and incubate at 37℃ for 1 hour in a roller drum (250 rpm) to allow cells to recover and express the antibiotic resistance marker. <br>
| |
| - | 15） Incubate on ice for 2 minutes. <br>
| |
| - | 16） Spread the appropriate quantity of cells (50 to 100 ml) on selective media. Store the remaining cells at 4℃. <br>
| |
| - | (A) E. coli cells from the control tube without DNA in step 12 above are plated on selective medium and nonselective medium. The first plating ensures that the selective medium is working properly since no growth should be observed. The second plating provides the number of viable cells in the absence of selective medium. <br>
| |
| - | (B) E. coli cells being tested for competency are plated on LB agar containing ampicillin (50 mg/ml final concentration) to ensure that the transformation efficiency has not decreased over time due to storage.<br>
| |
| - | 17） Incubate all plates overnight at 37℃ (agar side up). <br>
| |
| - | 18） Count the number of colonies.<br>
| |
| - | </p>
| |
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| - |
| |
| | var imageintro=new Array(data1,data2,data3); | | var imageintro=new Array(data1,data2,data3); |
| | var imagecount=0; | | var imagecount=0; |
"
