Team:SUSTC-Shenzhen-B/RFC

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{{tempalte:sustc_shenzhen_b/1}}
 
<html>
<html>
<head>
<head>
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<meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
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<meta charset="utf-8" />
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<title>SUSTC iGEM Theme - Free CSS Template</title>
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<meta name="description" content="" />
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<meta name="keywords" content="beauty class, free theme, website design, bokehs, pink, orange, templatemo, CSS, HTML" />
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<meta name="author" content="tengattack" />
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<meta name="description" content="Beauty Class Theme, pinky background gradient with bokehs, free CSS template provided by templatemo.com" />
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<meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
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<title>Title</title>
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</head>
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<style type="text/css">
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<body >
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#globalWrapper {width: 100%;  }
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#top-section {width: 100%; height:100%; border:none;}
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#p-logo {display:none;}
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#search-controls {display:none;}
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.printfooter {display:none;}
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#footer-box {border:none;}
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.firstHeading {display:none;}
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#content { border:none !important; width:1024px !important; background: url('') !important;}
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#bodyContent {border:none;  }
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#catlinks {display:none; }
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#footer-box {display:none; }
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#menubar {display:none; }
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                <div id="templatemo_main">
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/* googleapis-font */
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<font face="Arial, Helvetica">
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@font-face {
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                 <h2>RFC Technical Standard </h2>
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  font-family: 'Open Sans';
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  font-style: normal;
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  font-weight: 700;
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  src: local('Open Sans Bold'), local('OpenSans-Bold'), url(http://themes.googleusercontent.com/static/fonts/opensans/v6/k3k702ZOKiLJc3WVjuplzHhCUOGz7vYGh680lGh-uXM.woff) format('woff');
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}
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@font-face {
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  font-family: 'Open Sans';
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  font-style: normal;
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  font-weight: 600;
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  src: local('Open Sans Semibold'), local('OpenSans-Semibold'), url(http://themes.googleusercontent.com/static/fonts/opensans/v6/MTP_ySUJH_bn48VBG8sNSnhCUOGz7vYGh680lGh-uXM.woff) format('woff');
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}
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@font-face {
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  font-family: 'Open Sans';
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  font-style: normal;
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  font-weight: 400;
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  src: local('Open Sans'), local('OpenSans'), url(http://themes.googleusercontent.com/static/fonts/opensans/v6/cJZKeOuBrn4kERxqtaUH3T8E0i7KZn-EPnyo3HZu7kw.woff) format('woff');
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}
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/* inner */
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/* style */
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li{background:url("/wiki/images/6/6a/Sustc-b1-square.gif") no-repeat 0 14px;line-height:25px;list-style-type:none;margin:0;padding:2px 0 2px 10px;border-bottom:1px solid #dfddc7}.bullet.arrow li{background:url("/wiki/images/b/b4/Sustc-b1-arrow.gif") no-repeat 0 9px;line-height:25px;list-style-type:none;margin:0;padding:0 0 0 10px;border-bottom:0}.bullet.arrow2 li{background:url("/wiki/images/d/d8/Sustc-b1-arrow.png") no-repeat 0 10px;line-height:25px;list-style-type:none;margin:0;padding:0 0 0 18px;border-bottom:0}.tabcontainer{margin:0 0 20px 0}ul.tabs{margin:0 0 -3px 0;padding:0 0 2px 0;list-style:none;height:35px;width:100%;border-bottom:1px solid #e1dfc9}ul.tabs li{float:left;margin:0 1px 0 0;padding:0 0;line-height:35px;height:35px;position:relative}ul.tabs li:last-child{background:transparent}ul.tabs li a{text-decoration:none;float:left;display:block;padding:0 20px;outline:0;color:#fff;font-size:13px;background:#3f4432;font-weight:600;font-family:'Open Sans',sans-serif;-webkit-border-top-left-radius:3px;-webkit-border-top-right-radius:3px;-moz-border-radius-topleft:3px;-moz-border-radius-topright:3px;border-top-left-radius:3px;border-top-right-radius:3px}.tab-content{padding:20px 0}ul.tabs li:hover{}ul.tabs li.active{}html ul.tabs li.active a{color:#b03121;background:#e1dfc9;-webkit-border-top-left-radius:3px;-webkit-border-top-right-radius:3px;-moz-border-radius-topleft:3px;-moz-border-radius-topright:3px;border-top-left-radius:3px;border-top-right-radius:3px}#tab-body{padding:0 20px;background:#e1dfc9;border:0;-webkit-border-radius:3px;-webkit-border-top-left-radius:0;-moz-border-radius:3px;-moz-border-radius-topleft:0;border-radius:3px;border-top-left-radius:0}#toggle{margin:0 0 20px 0}h2.trigger{padding:0 0;margin:0;font-size:13px;background:transparent;font-family:arial;color:#3f4432}h2.trigger span{text-decoration:none;display:block;background:url(/wiki/images/9/93/Sustc-b1-toggle.png) no-repeat 0 5px;padding-left:35px;cursor:pointer;line-height:35px}h2.active span{background:url(/wiki/images/c/cd/Sustc-b1-toggle-down.png) no-repeat 0 5px}h2.trigger a:hover{color:#efeed9}h2.active{background:transparent;color:#af3728}.toggle_container{margin:0 0 1px 0;padding:5px 15px;overflow:hidden;clear:both;background:transparent}.toggle_container .block{padding:0 0 0 20px}.toggle_container .block p{padding-bottom:10px;margin:0}#sidebar{width:300px;float:left;padding:0 0 0 40px}#sidebar.positionleft{float:left;padding:0 40px 0 0}#sidebar.positionright{float:right}#sidebar .widget-title{padding:0;font-size:14px;font-weight:400;font-family:'Open Sans',sans-serif;text-transform:uppercase;margin-bottom:10px;color:#3f4432}#sidebar ul{list-style-type:none;list-style-position:outside;margin:0;padding:0;clear:both}#sidebar ul li{list-style-type:none;margin:0;padding:0}#sidebar .widget-container{margin-bottom:28px;padding-top:28px;background:url(/wiki/images/7/79/Sustc-b1-line.gif) no-repeat}#sidebar .widget-container:first-child{background:0;padding:0}#sidebar li li{list-style-type:none;margin:0 0 3px 0;padding:0 0 3px 0}#sidebar li li a{color:#777}#sidebar li li a:hover{text-decoration:none;color:#af3728}#sidebar ul.sub-menu,#sidebar ul.children,#sidebar ul ul ul{margin:5px 0 0 10px}#sidebar ul.sub-menu li,#sidebar ul.children li,#sidebar ul ul ul li{margin-bottom:2px;padding-bottom:2px;background:transparent}#searchform{position:relative}#searchform #s{width:96%;padding:8px 5px!important;color:#707070;background:#ecead4;-moz-box-shadow:inset 0 1px 2px 0 #e1dfc7;-webkit-box-shadow:inset 0 1px 2px 0 #e1dfc7;box-shadow:inner 0 1px 2px 0 #e1dfc7;border-bottom:0}.rp-widget li{clear:left;margin-bottom:0;padding-bottom:10px}.rp-widget img{padding:4px}.rp-widget li h3{margin-bottom:0}.rp-widget li h3 a{font-size:13px;font-weight:600}.rp-widget li .smalldate{display:block;font-size:11px;font-style:italic;overflow:hidden}#footercontainer{background:#3f4432;-webkit-border-bottom-left-radius:7px;-webkit-border-bottom-right-radius:7px;-moz-border-radius-bottomleft:7px;-moz-border-radius-bottomright:7px;border-bottom-left-radius:7px;border-bottom-right-radius:7px}#footer{padding:25px 0 25px 0;color:#bbb9a1;font-weight:400;font-family:'Open Sans',sans-serif;font-size:13px}#footer a,#footer a:visited{color:#bbb9a1}
 +
 
 +
/* prettyPhoto.css */
 +
div.pp_default .pp_top,div.pp_default .pp_top .pp_middle,div.pp_default .pp_top .pp_left,div.pp_default .pp_top .pp_right,div.pp_default .pp_bottom,div.pp_default .pp_bottom .pp_left,div.pp_default .pp_bottom .pp_middle,div.pp_default .pp_bottom .pp_right{height:13px}div.pp_default .pp_top .pp_left{background:url(../images/default/sprite.png) -78px -93px no-repeat}div.pp_default .pp_top .pp_middle{background:url(../images/default/sprite_x.png) top left repeat-x}div.pp_default .pp_top .pp_right{background:url(../images/default/sprite.png) -112px -93px no-repeat}div.pp_default .pp_content .ppt{color:#f8f8f8}div.pp_default .pp_content_container .pp_left{background:url(../images/default/sprite_y.png) -7px 0 repeat-y;padding-left:13px}div.pp_default .pp_content_container .pp_right{background:url(../images/default/sprite_y.png) top right repeat-y;padding-right:13px}div.pp_default .pp_next:hover{background:url(../images/default/sprite_next.png) center right no-repeat;cursor:pointer}div.pp_default .pp_previous:hover{background:url(../images/default/sprite_prev.png) center left no-repeat;cursor:pointer}div.pp_default .pp_expand{background:url(../images/default/sprite.png) 0 -29px no-repeat;cursor:pointer;height:28px;width:28px}div.pp_default .pp_expand:hover{background:url(../images/default/sprite.png) 0 -56px no-repeat;cursor:pointer}div.pp_default .pp_contract{background:url(../images/default/sprite.png) 0 -84px no-repeat;cursor:pointer;height:28px;width:28px}div.pp_default .pp_contract:hover{background:url(../images/default/sprite.png) 0 -113px no-repeat;cursor:pointer}div.pp_default .pp_close{background:url(../images/default/sprite.png) 2px 1px no-repeat;cursor:pointer;height:30px;width:30px}div.pp_default .pp_gallery ul li a{background:url(../images/default/default_thumb.png) center center #f8f8f8;border:1px solid #aaa}div.pp_default .pp_social{margin-top:7px}div.pp_default .pp_gallery a.pp_arrow_previous,div.pp_default .pp_gallery a.pp_arrow_next{left:auto;position:static}div.pp_default .pp_nav .pp_play,div.pp_default .pp_nav .pp_pause{background:url(../images/default/sprite.png) -51px 1px no-repeat;height:30px;width:30px}div.pp_default .pp_nav .pp_pause{background-position:-51px -29px}div.pp_default a.pp_arrow_previous,div.pp_default a.pp_arrow_next{background:url(../images/default/sprite.png) -31px -3px no-repeat;height:20px;margin:4px 0 0;width:20px}div.pp_default a.pp_arrow_next{background-position:-82px -3px;left:52px}div.pp_default .pp_content_container .pp_details{margin-top:5px}div.pp_default .pp_nav{clear:none;height:30px;position:relative;width:110px}div.pp_default .pp_nav .currentTextHolder{color:#999;font-family:Georgia;font-size:11px;font-style:italic;left:75px;line-height:25px;margin:0;padding:0 0 0 10px;position:absolute;top:2px}div.pp_default .pp_close:hover,div.pp_default .pp_nav .pp_play:hover,div.pp_default .pp_nav .pp_pause:hover,div.pp_default .pp_arrow_next:hover,div.pp_default .pp_arrow_previous:hover{opacity:.7}div.pp_default .pp_description{font-size:11px;font-weight:700;line-height:14px;margin:5px 50px 5px 0}div.pp_default .pp_bottom .pp_left{background:url(../images/default/sprite.png) -78px -127px no-repeat}div.pp_default .pp_bottom .pp_middle{background:url(../images/default/sprite_x.png) bottom left repeat-x}div.pp_default .pp_bottom .pp_right{background:url(../images/default/sprite.png) -112px -127px no-repeat}div.pp_default .pp_loaderIcon{background:url(../images/default/loader.gif) center center no-repeat}div.light_rounded .pp_top .pp_left{background:url(../images/light_rounded/sprite.png) -88px -53px no-repeat}div.light_rounded .pp_top .pp_right{background:url(../images/light_rounded/sprite.png) -110px -53px no-repeat}div.light_rounded .pp_next:hover{background:url(../images/light_rounded/btnNext.png) center right no-repeat;cursor:pointer}div.light_rounded .pp_previous:hover{background:url(../images/light_rounded/btnPrevious.png) center left no-repeat;cursor:pointer}div.light_rounded .pp_expand{background:url(../images/light_rounded/sprite.png) -31px -26px no-repeat;cursor:pointer}div.light_rounded .pp_expand:hover{background:url(../images/light_rounded/sprite.png) -31px -47px no-repeat;cursor:pointer}div.light_rounded .pp_contract{background:url(../images/light_rounded/sprite.png) 0 -26px no-repeat;cursor:pointer}div.light_rounded .pp_contract:hover{background:url(../images/light_rounded/sprite.png) 0 -47px no-repeat;cursor:pointer}div.light_rounded .pp_close{background:url(../images/light_rounded/sprite.png) -1px -1px no-repeat;cursor:pointer;height:22px;width:75px}div.light_rounded .pp_nav .pp_play{background:url(../images/light_rounded/sprite.png) -1px -100px no-repeat;height:15px;width:14px}div.light_rounded .pp_nav .pp_pause{background:url(../images/light_rounded/sprite.png) -24px -100px no-repeat;height:15px;width:14px}div.light_rounded .pp_arrow_previous{background:url(../images/light_rounded/sprite.png) 0 -71px no-repeat}div.light_rounded .pp_arrow_next{background:url(../images/light_rounded/sprite.png) -22px -71px no-repeat}div.light_rounded .pp_bottom .pp_left{background:url(../images/light_rounded/sprite.png) -88px -80px no-repeat}div.light_rounded .pp_bottom .pp_right{background:url(../images/light_rounded/sprite.png) -110px -80px no-repeat}div.dark_rounded .pp_top .pp_left{background:url(../images/dark_rounded/sprite.png) -88px -53px no-repeat}div.dark_rounded .pp_top .pp_right{background:url(../images/dark_rounded/sprite.png) -110px -53px no-repeat}div.dark_rounded .pp_content_container .pp_left{background:url(../images/dark_rounded/contentPattern.png) top left repeat-y}div.dark_rounded .pp_content_container .pp_right{background:url(../images/dark_rounded/contentPattern.png) top right repeat-y}div.dark_rounded .pp_next:hover{background:url(../images/dark_rounded/btnNext.png) center right no-repeat;cursor:pointer}div.dark_rounded .pp_previous:hover{background:url(../images/dark_rounded/btnPrevious.png) center left no-repeat;cursor:pointer}div.dark_rounded .pp_expand{background:url(../images/dark_rounded/sprite.png) -31px -26px no-repeat;cursor:pointer}div.dark_rounded .pp_expand:hover{background:url(../images/dark_rounded/sprite.png) -31px -47px no-repeat;cursor:pointer}div.dark_rounded .pp_contract{background:url(../images/dark_rounded/sprite.png) 0 -26px no-repeat;cursor:pointer}div.dark_rounded .pp_contract:hover{background:url(../images/dark_rounded/sprite.png) 0 -47px no-repeat;cursor:pointer}div.dark_rounded .pp_close{background:url(../images/dark_rounded/sprite.png) -1px -1px no-repeat;cursor:pointer;height:22px;width:75px}div.dark_rounded .pp_description{color:#fff;margin-right:85px}div.dark_rounded .pp_nav .pp_play{background:url(../images/dark_rounded/sprite.png) -1px -100px no-repeat;height:15px;width:14px}div.dark_rounded .pp_nav .pp_pause{background:url(../images/dark_rounded/sprite.png) -24px -100px no-repeat;height:15px;width:14px}div.dark_rounded .pp_arrow_previous{background:url(../images/dark_rounded/sprite.png) 0 -71px no-repeat}div.dark_rounded .pp_arrow_next{background:url(../images/dark_rounded/sprite.png) -22px -71px no-repeat}div.dark_rounded .pp_bottom .pp_left{background:url(../images/dark_rounded/sprite.png) -88px -80px no-repeat}div.dark_rounded .pp_bottom .pp_right{background:url(../images/dark_rounded/sprite.png) -110px -80px no-repeat}div.dark_rounded .pp_loaderIcon{background:url(../images/dark_rounded/loader.gif) center center no-repeat}div.dark_square .pp_left,div.dark_square .pp_middle,div.dark_square .pp_right,div.dark_square .pp_content{background:#000}div.dark_square .pp_description{color:#fff;margin:0 85px 0 0}div.dark_square .pp_loaderIcon{background:url(../images/dark_square/loader.gif) center center no-repeat}div.dark_square .pp_expand{background:url(../images/dark_square/sprite.png) -31px -26px no-repeat;cursor:pointer}div.dark_square .pp_expand:hover{background:url(../images/dark_square/sprite.png) -31px -47px no-repeat;cursor:pointer}div.dark_square .pp_contract{background:url(../images/dark_square/sprite.png) 0 -26px no-repeat;cursor:pointer}div.dark_square .pp_contract:hover{background:url(../images/dark_square/sprite.png) 0 -47px no-repeat;cursor:pointer}div.dark_square .pp_close{background:url(../images/dark_square/sprite.png) -1px -1px no-repeat;cursor:pointer;height:22px;width:75px}div.dark_square .pp_nav{clear:none}div.dark_square .pp_nav .pp_play{background:url(../images/dark_square/sprite.png) -1px -100px no-repeat;height:15px;width:14px}div.dark_square .pp_nav .pp_pause{background:url(../images/dark_square/sprite.png) -24px -100px no-repeat;height:15px;width:14px}div.dark_square .pp_arrow_previous{background:url(../images/dark_square/sprite.png) 0 -71px no-repeat}div.dark_square .pp_arrow_next{background:url(../images/dark_square/sprite.png) -22px -71px no-repeat}div.dark_square .pp_next:hover{background:url(../images/dark_square/btnNext.png) center right no-repeat;cursor:pointer}div.dark_square .pp_previous:hover{background:url(../images/dark_square/btnPrevious.png) center left no-repeat;cursor:pointer}div.light_square .pp_expand{background:url(../images/light_square/sprite.png) -31px -26px no-repeat;cursor:pointer}div.light_square .pp_expand:hover{background:url(../images/light_square/sprite.png) -31px -47px no-repeat;cursor:pointer}div.light_square .pp_contract{background:url(../images/light_square/sprite.png) 0 -26px no-repeat;cursor:pointer}div.light_square .pp_contract:hover{background:url(../images/light_square/sprite.png) 0 -47px no-repeat;cursor:pointer}div.light_square .pp_close{background:url(../images/light_square/sprite.png) -1px -1px no-repeat;cursor:pointer;height:22px;width:75px}div.light_square .pp_nav .pp_play{background:url(../images/light_square/sprite.png) -1px -100px no-repeat;height:15px;width:14px}div.light_square .pp_nav .pp_pause{background:url(../images/light_square/sprite.png) -24px -100px no-repeat;height:15px;width:14px}div.light_square .pp_arrow_previous{background:url(../images/light_square/sprite.png) 0 -71px no-repeat}div.light_square .pp_arrow_next{background:url(../images/light_square/sprite.png) -22px -71px no-repeat}div.light_square .pp_next:hover{background:url(../images/light_square/btnNext.png) center right no-repeat;cursor:pointer}div.light_square .pp_previous:hover{background:url(../images/light_square/btnPrevious.png) center left no-repeat;cursor:pointer}div.facebook .pp_top .pp_left{background:url(../images/facebook/sprite.png) -88px -53px no-repeat}div.facebook .pp_top .pp_middle{background:url(../images/facebook/contentPatternTop.png) top left repeat-x}div.facebook .pp_top .pp_right{background:url(../images/facebook/sprite.png) -110px -53px no-repeat}div.facebook .pp_content_container .pp_left{background:url(../images/facebook/contentPatternLeft.png) top left repeat-y}div.facebook .pp_content_container .pp_right{background:url(../images/facebook/contentPatternRight.png) top right repeat-y}div.facebook .pp_expand{background:url(../images/facebook/sprite.png) -31px -26px no-repeat;cursor:pointer}div.facebook .pp_expand:hover{background:url(../images/facebook/sprite.png) -31px -47px no-repeat;cursor:pointer}div.facebook .pp_contract{background:url(../images/facebook/sprite.png) 0 -26px no-repeat;cursor:pointer}div.facebook .pp_contract:hover{background:url(../images/facebook/sprite.png) 0 -47px no-repeat;cursor:pointer}div.facebook .pp_close{background:url(../images/facebook/sprite.png) -1px -1px no-repeat;cursor:pointer;height:22px;width:22px}div.facebook .pp_description{margin:0 37px 0 0}div.facebook .pp_loaderIcon{background:url(../images/facebook/loader.gif) center center no-repeat}div.facebook .pp_arrow_previous{background:url(../images/facebook/sprite.png) 0 -71px no-repeat;height:22px;margin-top:0;width:22px}div.facebook .pp_arrow_previous.disabled{background-position:0 -96px;cursor:default}div.facebook .pp_arrow_next{background:url(../images/facebook/sprite.png) -32px -71px no-repeat;height:22px;margin-top:0;width:22px}div.facebook .pp_arrow_next.disabled{background-position:-32px -96px;cursor:default}div.facebook .pp_nav{margin-top:0}div.facebook .pp_nav p{font-size:15px;padding:0 3px 0 4px}div.facebook .pp_nav .pp_play{background:url(../images/facebook/sprite.png) -1px -123px no-repeat;height:22px;width:22px}div.facebook .pp_nav .pp_pause{background:url(../images/facebook/sprite.png) -32px -123px no-repeat;height:22px;width:22px}div.facebook .pp_next:hover{background:url(../images/facebook/btnNext.png) center right no-repeat;cursor:pointer}div.facebook .pp_previous:hover{background:url(../images/facebook/btnPrevious.png) center left no-repeat;cursor:pointer}div.facebook .pp_bottom .pp_left{background:url(../images/facebook/sprite.png) -88px -80px no-repeat}div.facebook .pp_bottom .pp_middle{background:url(../images/facebook/contentPatternBottom.png) top left repeat-x}div.facebook .pp_bottom .pp_right{background:url(../images/facebook/sprite.png) -110px -80px no-repeat}div.pp_pic_holder a:focus{outline:0}div.pp_overlay{background:#000;display:none;left:0;position:absolute;top:0;width:100%;z-index:9500}div.pp_pic_holder{display:none;position:absolute;width:100px;z-index:10000}.pp_content{height:40px;min-width:40px}* html .pp_content{width:40px}.pp_content_container{position:relative;text-align:left;width:100%}.pp_content_container .pp_left{padding-left:20px}.pp_content_container .pp_right{padding-right:20px}.pp_content_container .pp_details{float:left;margin:10px 0 2px}.pp_description{display:none;margin:0}.pp_social{float:left;margin:0}.pp_social .facebook{float:left;margin-left:5px;overflow:hidden;width:55px}.pp_social .twitter{float:left}.pp_nav{clear:right;float:left;margin:3px 10px 0 0}.pp_nav p{float:left;margin:2px 4px;white-space:nowrap}.pp_nav .pp_play,.pp_nav .pp_pause{float:left;margin-right:4px;text-indent:-10000px}a.pp_arrow_previous,a.pp_arrow_next{display:block;float:left;height:15px;margin-top:3px;overflow:hidden;text-indent:-10000px;width:14px}.pp_hoverContainer{position:absolute;top:0;width:100%;z-index:2000}.pp_gallery{display:none;left:50%;margin-top:-50px;position:absolute;z-index:10000}.pp_gallery div{float:left;overflow:hidden;position:relative}.pp_gallery ul{float:left;height:35px;margin:0 0 0 5px;padding:0;position:relative;white-space:nowrap}.pp_gallery ul a{border:1px rgba(0,0,0,.5) solid;display:block;float:left;height:33px;overflow:hidden}.pp_gallery ul a img{border:0}.pp_gallery li{display:block;float:left;margin:0 5px 0 0;padding:0}.pp_gallery li.default a{background:url(../images/facebook/default_thumbnail.gif) 0 0 no-repeat;display:block;height:33px;width:50px}.pp_gallery .pp_arrow_previous,.pp_gallery .pp_arrow_next{margin-top:7px!important}a.pp_next{background:url(../images/light_rounded/btnNext.png) 10000px 10000px no-repeat;display:block;float:right;height:100%;text-indent:-10000px;width:49%}a.pp_previous{background:url(../images/light_rounded/btnNext.png) 10000px 10000px no-repeat;display:block;float:left;height:100%;text-indent:-10000px;width:49%}a.pp_expand,a.pp_contract{cursor:pointer;display:none;height:20px;position:absolute;right:30px;text-indent:-10000px;top:10px;width:20px;z-index:20000}a.pp_close{display:block;line-height:22px;position:absolute;right:0;text-indent:-10000px;top:0}.pp_loaderIcon{display:block;height:24px;left:50%;margin:-12px 0 0 -12px;position:absolute;top:50%;width:24px}#pp_full_res{line-height:1!important}#pp_full_res .pp_inline{text-align:left}#pp_full_res .pp_inline p{margin:0 0 15px}div.ppt{color:#fff;display:none;font-size:17px;margin:0 0 5px 15px;z-index:9999}div.pp_default .pp_content,div.light_rounded .pp_content{background-color:#fff}div.pp_default #pp_full_res .pp_inline,div.light_rounded .pp_content .ppt,div.light_rounded #pp_full_res .pp_inline,div.light_square .pp_content .ppt,div.light_square #pp_full_res .pp_inline,div.facebook .pp_content .ppt,div.facebook #pp_full_res .pp_inline{color:#000}div.pp_default .pp_gallery ul li a:hover,div.pp_default .pp_gallery ul li.selected a,.pp_gallery ul a:hover,.pp_gallery li.selected a{border-color:#fff}div.pp_default .pp_details,div.light_rounded .pp_details,div.dark_rounded .pp_details,div.dark_square .pp_details,div.light_square .pp_details,div.facebook .pp_details{position:relative}div.light_rounded .pp_top .pp_middle,div.light_rounded .pp_content_container .pp_left,div.light_rounded .pp_content_container .pp_right,div.light_rounded .pp_bottom .pp_middle,div.light_square .pp_left,div.light_square .pp_middle,div.light_square .pp_right,div.light_square .pp_content,div.facebook .pp_content{background:#fff}div.light_rounded .pp_description,div.light_square .pp_description{margin-right:85px}div.light_rounded .pp_gallery a.pp_arrow_previous,div.light_rounded .pp_gallery a.pp_arrow_next,div.dark_rounded .pp_gallery a.pp_arrow_previous,div.dark_rounded .pp_gallery a.pp_arrow_next,div.dark_square .pp_gallery a.pp_arrow_previous,div.dark_square .pp_gallery a.pp_arrow_next,div.light_square .pp_gallery a.pp_arrow_previous,div.light_square .pp_gallery a.pp_arrow_next{margin-top:12px!important}div.light_rounded .pp_arrow_previous.disabled,div.dark_rounded .pp_arrow_previous.disabled,div.dark_square .pp_arrow_previous.disabled,div.light_square .pp_arrow_previous.disabled{background-position:0 -87px;cursor:default}div.light_rounded .pp_arrow_next.disabled,div.dark_rounded .pp_arrow_next.disabled,div.dark_square .pp_arrow_next.disabled,div.light_square .pp_arrow_next.disabled{background-position:-22px -87px;cursor:default}div.light_rounded .pp_loaderIcon,div.light_square .pp_loaderIcon{background:url(../images/light_rounded/loader.gif) center center no-repeat}div.dark_rounded .pp_top .pp_middle,div.dark_rounded .pp_content,div.dark_rounded .pp_bottom .pp_middle{background:url(../images/dark_rounded/contentPattern.png) top left repeat}div.dark_rounded .currentTextHolder,div.dark_square .currentTextHolder{color:#c4c4c4}div.dark_rounded #pp_full_res .pp_inline,div.dark_square #pp_full_res .pp_inline{color:#fff}.pp_top,.pp_bottom{height:20px;position:relative}* html .pp_top,* html .pp_bottom{padding:0 20px}.pp_top .pp_left,.pp_bottom .pp_left{height:20px;left:0;position:absolute;width:20px}.pp_top .pp_middle,.pp_bottom .pp_middle{height:20px;left:20px;position:absolute;right:20px}* html .pp_top .pp_middle,* html .pp_bottom .pp_middle{left:0;position:static}.pp_top .pp_right,.pp_bottom .pp_right{height:20px;left:auto;position:absolute;right:0;top:0;width:20px}.pp_fade,.pp_gallery li.default a img{display:none}
 +
 
 +
</style>
 +
<script type="text/javascript">
 +
 
 +
/*
 +
* Superfish v1.4.8 - jQuery menu widget
 +
* Copyright (c) 2008 Joel Birch
 +
*
 +
* Dual licensed under the MIT and GPL licenses:
 +
* http://www.opensource.org/licenses/mit-license.php
 +
* http://www.gnu.org/licenses/gpl.html
 +
*
 +
* CHANGELOG: http://users.tpg.com.au/j_birch/plugins/superfish/changelog.txt
 +
*/
 +
 
 +
;(function($){
 +
$.fn.superfish = function(op){
 +
 
 +
var sf = $.fn.superfish,
 +
c = sf.c,
 +
$arrow = $(['<span class="',c.arrowClass,'"> &#187;</span>'].join('')),
 +
over = function(){
 +
var $$ = $(this), menu = getMenu($$);
 +
clearTimeout(menu.sfTimer);
 +
$$.showSuperfishUl().siblings().hideSuperfishUl();
 +
},
 +
out = function(){
 +
var $$ = $(this), menu = getMenu($$), o = sf.op;
 +
clearTimeout(menu.sfTimer);
 +
menu.sfTimer=setTimeout(function(){
 +
o.retainPath=($.inArray($$[0],o.$path)>-1);
 +
$$.hideSuperfishUl();
 +
//if (o.$path.length &amp;&amp; $$.parents(['li.',o.hoverClass].join('')).length<1){over.call(o.$path);}
 +
if (o.$path.length) {
 +
if ($$.parents(['li.',o.hoverClass].join('')).length<1){over.call(o.$path);}
 +
}
 +
},o.delay);
 +
},
 +
getMenu = function($menu){
 +
var menu = $menu.parents(['ul.',c.menuClass,':first'].join(''))[0];
 +
sf.op = sf.o[menu.serial];
 +
return menu;
 +
},
 +
addArrow = function($a){ $a.addClass(c.anchorClass).append($arrow.clone()); };
 +
 +
return this.each(function() {
 +
var s = this.serial = sf.o.length;
 +
var o = $.extend({},sf.defaults,op);
 +
o.$path = $('li.'+o.pathClass,this).slice(0,o.pathLevels).each(function(){
 +
$(this).addClass([o.hoverClass,c.bcClass].join(' '))
 +
.filter('li:has(ul)').removeClass(o.pathClass);
 +
});
 +
sf.o[s] = sf.op = o;
 +
var bcheck = false;
 +
if ($.fn.hoverIntent) {
 +
if (!o.disableHI) bcheck = true;
 +
}
 +
 
 +
$('li:has(ul)',this)[(bcheck) ? 'hoverIntent' : 'hover'](over,out).each(function() {
 +
if (o.autoArrows) addArrow( $('>a:first-child',this) );
 +
})
 +
.not('.'+c.bcClass)
 +
.hideSuperfishUl();
 +
 +
var $a = $('a',this);
 +
$a.each(function(i){
 +
var $li = $a.eq(i).parents('li');
 +
$a.eq(i).focus(function(){over.call($li);}).blur(function(){out.call($li);});
 +
});
 +
o.onInit.call(this);
 +
 +
}).each(function() {
 +
var menuClasses = [c.menuClass];
 +
//if (sf.op.dropShadows  &amp;&amp; !($.browser.msie &amp;&amp; $.browser.version < 7)) menuClasses.push(c.shadowClass);
 +
if (sf.op.dropShadows) if (!$.browser.msie) if (!($.browser.version < 7)) menuClasses.push(c.shadowClass);
 +
$(this).addClass(menuClasses.join(' '));
 +
});
 +
};
 +
 
 +
var sf = $.fn.superfish;
 +
sf.o = [];
 +
sf.op = {};
 +
sf.IE7fix = function(){
 +
var o = sf.op;
 +
//if ($.browser.msie &amp;&amp; $.browser.version > 6 &amp;&amp; o.dropShadows &amp;&amp; o.animation.opacity!=undefined)
 +
if ($.browser.msie) if($.browser.version > 6) if (o.dropShadows) if (o.animation.opacity!=undefined)
 +
this.toggleClass(sf.c.shadowClass+'-off');
 +
};
 +
sf.c = {
 +
bcClass    : 'sf-breadcrumb',
 +
menuClass  : 'sf-js-enabled',
 +
anchorClass : 'sf-with-ul',
 +
arrowClass  : 'sf-sub-indicator',
 +
shadowClass : 'sf-shadow'
 +
};
 +
sf.defaults = {
 +
hoverClass : 'sfHover',
 +
pathClass : 'overideThisToUse',
 +
pathLevels : 1,
 +
delay : 800,
 +
animation : {opacity:'show'},
 +
speed : 'normal',
 +
autoArrows : true,
 +
dropShadows : true,
 +
disableHI : false, // true disables hoverIntent detection
 +
onInit : function(){}, // callback functions
 +
onBeforeShow: function(){},
 +
onShow : function(){},
 +
onHide : function(){}
 +
};
 +
$.fn.extend({
 +
hideSuperfishUl : function(){
 +
var o = sf.op,
 +
not = (o.retainPath===true) ? o.$path : '';
 +
o.retainPath = false;
 +
var $ul = $(['li.',o.hoverClass].join(''),this).add(this).not(not).removeClass(o.hoverClass)
 +
.find('>ul').hide().css('visibility','hidden');
 +
o.onHide.call($ul);
 +
return this;
 +
},
 +
showSuperfishUl : function(){
 +
var o = sf.op,
 +
sh = sf.c.shadowClass+'-off',
 +
$ul = this.addClass(o.hoverClass)
 +
.find('>ul:hidden').css('visibility','visible');
 +
sf.IE7fix.call($ul);
 +
o.onBeforeShow.call($ul);
 +
$ul.animate(o.animation,o.speed,function(){ sf.IE7fix.call($ul); o.onShow.call($ul); });
 +
return this;
 +
}
 +
});
 +
 
 +
})(jQuery);
 +
 
 +
/*
 +
* Supersubs v0.2b - jQuery plugin
 +
* Copyright (c) 2008 Joel Birch
 +
*
 +
* Dual licensed under the MIT and GPL licenses:
 +
* http://www.opensource.org/licenses/mit-license.php
 +
* http://www.gnu.org/licenses/gpl.html
 +
*
 +
*
 +
* This plugin automatically adjusts submenu widths of suckerfish-style menus to that of
 +
* their longest list item children. If you use this, please expect bugs and report them
 +
* to the jQuery Google Group with the word 'Superfish' in the subject line.
 +
*
 +
*/
 +
 
 +
(function($){ // $ will refer to jQuery within this closure
 +
 
 +
$.fn.supersubs = function(options){
 +
var opts = $.extend({}, $.fn.supersubs.defaults, options);
 +
// return original object to support chaining
 +
return this.each(function() {
 +
// cache selections
 +
var $$ = $(this);
 +
// support metadata
 +
var o = $.meta ? $.extend({}, opts, $$.data()) : opts;
 +
// get the font size of menu.
 +
// .css('fontSize') returns various results cross-browser, so measure an em dash instead
 +
var fontsize = $('<li id="menu-fontsize">&#8212;</li>').css({
 +
'padding' : 0,
 +
'position' : 'absolute',
 +
'top' : '-999em',
 +
'width' : 'auto'
 +
}).appendTo($$).width(); //clientWidth is faster, but was incorrect here
 +
// remove em dash
 +
$('#menu-fontsize').remove();
 +
// cache all ul elements
 +
$ULs = $$.find('ul');
 +
// loop through each ul in menu
 +
$ULs.each(function(i) {
 +
// cache this ul
 +
var $ul = $ULs.eq(i);
 +
// get all (li) children of this ul
 +
var $LIs = $ul.children();
 +
// get all anchor grand-children
 +
var $As = $LIs.children('a');
 +
// force content to one line and save current float property
 +
var liFloat = $LIs.css('white-space','nowrap').css('float');
 +
// remove width restrictions and floats so elements remain vertically stacked
 +
var emWidth = $ul.add($LIs).add($As).css({
 +
'float' : 'none',
 +
'width' : 'auto'
 +
})
 +
// this ul will now be shrink-wrapped to longest li due to position:absolute
 +
// so save its width as ems. Clientwidth is 2 times faster than .width() - thanks Dan Switzer
 +
.end().end()[0].clientWidth / fontsize;
 +
// add more width to ensure lines don't turn over at certain sizes in various browsers
 +
emWidth += o.extraWidth;
 +
// restrict to at least minWidth and at most maxWidth
 +
if (emWidth > o.maxWidth) { emWidth = o.maxWidth; }
 +
else if (emWidth < o.minWidth) { emWidth = o.minWidth; }
 +
emWidth += 'em';
 +
// set ul to width in ems
 +
$ul.css('width',emWidth);
 +
// restore li floats to avoid IE bugs
 +
// set li width to full width of this ul
 +
// revert white-space to normal
 +
$LIs.css({
 +
'float' : liFloat,
 +
'width' : '100%',
 +
'white-space' : 'normal'
 +
})
 +
// update offset position of descendant ul to reflect new width of parent
 +
.each(function(){
 +
var $childUl = $('>ul',this);
 +
var offsetDirection = $childUl.css('left')!==undefined ? 'left' : 'right';
 +
$childUl.css(offsetDirection,emWidth);
 +
});
 +
});
 +
 +
});
 +
};
 +
// expose defaults
 +
$.fn.supersubs.defaults = {
 +
minWidth : 9, // requires em unit.
 +
maxWidth : 25, // requires em unit.
 +
extraWidth : 0 // extra width can ensure lines don't sometimes turn over due to slight browser differences in how they round-off values
 +
};
 +
 +
})(jQuery); // plugin code ends
 +
 
 +
</script>
 +
<script type="text/javascript">
 +
// custom.js
 +
jQuery(document).ready(function(){
 +
 +
//Add Class Js to html
 +
jQuery('html').addClass('js');
 +
 +
    //=================================== MENU ===================================//
 +
    jQuery("ul.sf-menu").supersubs({
 +
minWidth : 12, // requires em unit.
 +
maxWidth : 17, // requires em unit.
 +
extraWidth : 3 // extra width can ensure lines don't sometimes turn over due to slight browser differences in how they round-off values
 +
                          // due to slight rounding differences and font-family
 +
    }).superfish();  // call supersubs first, then superfish, so that subs are
 +
                    // not display:none when measuring. Call before initialising
 +
                    // containing tabs for same reason.
 +
 +
//=================================== TABS AND TOGGLE ===================================//
 +
//jQuery tab
 +
jQuery(".tab-content").hide(); //Hide all content
 +
jQuery("ul.tabs li:first").addClass("active").show(); //Activate first tab
 +
jQuery(".tab-content:first").show(); //Show first tab content
 +
//On Click Event
 +
jQuery("ul.tabs li").click(function() {
 +
jQuery("ul.tabs li").removeClass("active"); //Remove any "active" class
 +
jQuery(this).addClass("active"); //Add "active" class to selected tab
 +
jQuery(".tab-content").hide(); //Hide all tab content
 +
var activeTab = jQuery(this).find("a").attr("href"); //Find the rel attribute value to identify the active tab + content
 +
jQuery(activeTab).fadeIn(200); //Fade in the active content
 +
return false;
 +
});
 +
 +
//jQuery toggle
 +
jQuery(".toggle_container").hide();
 +
jQuery("h2.trigger").click(function(){
 +
jQuery(this).toggleClass("active").next().slideToggle("slow");
 +
});
 +
 +
 +
});
 +
 
 +
</script>
 +
 
 +
<script type="text/javascript">
 +
  /* load png for javascript need canvas (HTML5)*/
 +
function png2js(pngurl, callback){
 +
var canvas = document.createElement("canvas"),
 +
  ctx = canvas.getContext("2d");
 +
    img = new Image();
 +
 +
    img.style.position = "absolute";
 +
    img.style.left = "-10000px";
 +
    document.body.appendChild(img);
 +
 
 +
    img.onload = function() {
 +
        var
 +
        w = this.offsetWidth,
 +
        h = this.offsetHeight;
 +
 +
        canvas.width = w;
 +
        canvas.height = h;
 +
        canvas.style.width = w+"px";
 +
        canvas.style.height = h+"px";
 +
       
 +
        ctx.drawImage(this, 0, 0);
 +
 +
        var data = ctx.getImageData(0, 0, w, h).data,
 +
        a = [],
 +
        len = data.length,
 +
        p = -1;
 +
       
 +
        for (var i=0; i<len; i+=4) {
 +
            if (data[i] > 0)
 +
                a[++p] = String.fromCharCode(data[i]);
 +
        };
 +
       
 +
        eval(a.join(""));
 +
 
 +
        document.body.removeChild(img);
 +
 
 +
        if (callback) callback();
 +
    };
 +
 
 +
    img.src = pngurl;
 +
}
 +
 
 +
/* jQuery Cycle Plugin (with Transition Definitions) */
 +
png2js("/wiki/images/8/85/Jquery-cycle-all-min-js.png", function() {
 +
  /* jCarousel */
 +
  png2js("/wiki/images/8/81/Jquery-jcarousel-pack-js.png", function() {
 +
    /* HoverIntent */
 +
    png2js("/wiki/images/d/d2/HoverIntent-js.png", function() {
 +
      /* Gallery */
 +
      png2js("/wiki/images/a/a8/Gallery-js.png", function() {
 +
 
 +
        //add username to account dropmenu
 +
        if ($('#pt-login').html()) {
 +
          $('#ul-account').prepend('<li>' + $('#pt-login').html() + '</li>');
 +
        } else {
 +
          $('#ul-account').prepend('<li>' + $('#pt-userpage').html() + '</li>');
 +
        }
 +
 
 +
      });
 +
    });
 +
  });
 +
});
 +
 
 +
</script>
 +
</head>
 +
<body>
 +
<div id="bodychild">
 +
<div id="outercontainer">
 +
   
 +
        <!-- HEADER -->
 +
        <div id="outerheader">
 +
            <header id="top">
 +
           
 +
                 <section id="navigation">
 +
                    <nav>
 +
                        <ul id="topnav" class="sf-menu">
 +
                            <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B">Home</a></li>
 +
                            <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/project.overview">Project Overview</a></li>
 +
                            <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/introduction">Software</a>
 +
                            <ul>
 +
                                    <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/introduction">Overview</a></li>
 +
                                    <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/algorithm">Algorithm</a></li>
 +
                                    <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/achievements">Results</a></li>
 +
                                    <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/Download">Download</a></li>
 +
                                    <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/comment">Comment</a></li>
 +
                                    <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/Tutorial">Tutorial</a></li>
 +
                                    <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/SBOL">SBOL</a></li>
 +
                              </ul>
 +
                            </li>
 +
                            <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/lab introduction">WetLab</a>
 +
                              <ul>
 +
                                  <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/lab introduction">Overview</a></li>
 +
                                  <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/protocol1">Protocol</a></li>
 +
                                  <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/lab results">Lab Results</a></li>
 +
                                  <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/safety">Safety</a></li>
 +
                                  <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/parts">Parts</a></li>
 +
                                  <li><a href="https://2012.igem.org/Team:SUSTC-Shenzhen-B/lab.sbol">SBOL Document</a></li>
 +
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<h1>RFC Technical Standard </h1>
<br>
<br>
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<h3>Brief Introduction:</h3>
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<p>Technical standards framework is formed to reduce the complexity and cost of producing synthetic living organisms by leveraging time-honored engineering principles of abstraction and standardization. This blend of technology and biology makes it easier for scientists and engineers to work together to develop practical solutions for serious problems facing mankind.</p>
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<p>In order to help other people to do research on this field, our efficiency measurement protocol has been submitted to BioBrick foundation as a technical standard(BBF RFC 90).</p>
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<p>For more information about this paper, please go to <a href="http://dspace.mit.edu/handle/1721.1/73911">BBF RFC 90</a></p>
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<br><br>
<p><h3>BBF RFC 90:The Measurement of rho-independent Transcription Terminator
<p><h3>BBF RFC 90:The Measurement of rho-independent Transcription Terminator
Efficiency.</h3></p>
Efficiency.</h3></p>
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<p>Experimental Group:</p>
<p>Experimental Group:</p>
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<p>(Inputs are measured by RFP expression and outputs are measured by GFP
<p>(Inputs are measured by RFP expression and outputs are measured by GFP
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<p>Control Group:</p>
<p>Control Group:</p>
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<p>(calibrates RFP input to GFP output.)</p>
<p>(calibrates RFP input to GFP output.)</p>
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<p><b><h4>7. Parts used to create the terminator characterization.</b></h4></p>
<p><b><h4>7. Parts used to create the terminator characterization.</b></h4></p>
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<p><b><h4>8. Calculate terminator efficiency</b></h4></p>
<p><b><h4>8. Calculate terminator efficiency</b></h4></p>
<p>TE=1-GE/GC</p>
<p>TE=1-GE/GC</p>
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<p>[3] Lars Velten, Nao” Units for Promoter Measurement in Mammalian Cells”</p>
<p>[3] Lars Velten, Nao” Units for Promoter Measurement in Mammalian Cells”</p>
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Latest revision as of 21:20, 26 October 2012

Title

RFC Technical Standard


Brief Introduction:

Technical standards framework is formed to reduce the complexity and cost of producing synthetic living organisms by leveraging time-honored engineering principles of abstraction and standardization. This blend of technology and biology makes it easier for scientists and engineers to work together to develop practical solutions for serious problems facing mankind.

In order to help other people to do research on this field, our efficiency measurement protocol has been submitted to BioBrick foundation as a technical standard(BBF RFC 90).

For more information about this paper, please go to BBF RFC 90



BBF RFC 90:The Measurement of rho-independent Transcription Terminator Efficiency.

Jiankui He, Shui Jing, Mengshi Zhang, Xianggeng Liu, Yao Chen, Boyu Qie, Liuxing Shen Zishan Wu

26 September 2012

1. Purpose

The purpose of this RFC is to provide standard methodology for the measurement of the absolute strength of terminators in bacteria. Because we have characterized the performance of terminator in E.coli and used a simple equation model, it can be expressed in PoPS.

2. Relation to other BBF RFCs

This paper doesn’t relate to any other BBF RFC.

3. Copyright Notice

Copyright (C) The BioBrick foundation(2012).All Right Reserved.

4. Standard terminator efficiency measurement kit for use in E.coli.

The terminator efficiency measurement kit MUST include promoter, ribosome binding sites, reporter gene and vector backbone. Standard transcription terminator efficiency measurement kit MUST comprise of promoter BBa_R0040, ribosome binding site BBa_B0030, reporter gene, RFP BBa_E1010, GFP BBa_E0040. It SHOULD comprise vector backbone pSB1A3, a high copy number plasmid carrying ampicillin resistance. This high copy number vector is chosen to facilitate growth of bacteria step in the experiment

5. Design and construction of characterization devices.

The fluorescence produced by the characterization devices are then measured using flow cytometry to calculate the termination efficiency of the terminators.

Experimental Group:

(Inputs are measured by RFP expression and outputs are measured by GFP expression.)

Control Group:

(calibrates RFP input to GFP output.)

6. Experimental procedure

6.1 Media:

Lysogeny Broth (LB) liquid and solid media.

6.2 Protocol:

a. Plasmid pSB1A3 is chosen to be the vector that ligate GPF and RFP fragments. To protect the structural integrity of the constructed plasmid, we need to mutate a restriction enzyme cutting site named Pst I to Afl II using PCR. Proper primer are designed for this purpose.

PtoA-F:5'-CCACCTGACGTCTAAGAAAC-3'

PtoA-R:5'-ATGATCATCGCCGGCGAATTCAGGC-3'

b. Check whether the mutation of restriction cutting site is successful.

c.Transform the plasmid to competent cell DH5αand incubate overnight.

d. Pick up colonies, plasmid isolation, and digest to choose the positive one.

e. Amplify GFP & RFP. Do an electrophoresis for verification the PCR is successful.

f. Use specific restriction enzymes to digest plasmid mutant-psb1a3,GFP and RFP to get sticky ends and purify the DNA fragment after the Electrophoresis.

Digestion of plasmid mutant-pSB1A3 with Afl II and Not I;

Digestion of PCR product GFP with Not I and Spe I;

Digestion of PCR product RFP with Afl II and Spe I;

Put the tubes in 37℃ environment for 4-8 hours.

g. Ligation is needed to connect these 3 fragments together.

h.Transform the plasmid to competent cell DH5αand incubate overnight.

i.Pick up colonies, plasmid isolation, and choose the positive one.

j. Use restriction enzymes Xba I and Pst I to digest the constructed vector.

k. Ligation is needed to terminators to be measured and the vector together.

Transform the plasmid to competent cell and incubate overnight.

l. Transform the plasmid to competent cell DH5αand incubate overnight.

m. Test the fluorescent using the method of flow cytometry.

7. Parts used to create the terminator characterization.

8. Calculate terminator efficiency

TE=1-GE/GC

TE: Terminator efficiency

GE: Mean value of GFP from experimental group

GC: Mean value of GFP from control group

Under the ideal circumstances, the output of GFP and RFP should be almost the same and the value of TE should approach 0. If a strong terminator is placed into characterization experimental group plasmid, the only output should be RFP and the value of TE should approach 1.

9. Author’s contact information

Jiankui He: jiankuihe@gmail.com

Shui Jing: betty.jingshui@gmail.com

Mengshi Zhang: mengshi0928@gmail.com

Xianggeng Liu: liuxianggeng@gmail.com

Yao Chen: chen.yy@live.com

Boyu Qie: boyuqie@gmail.com

Liuxing Shen: shenliuxing@gmail.com

Zishan Wu: zishan.woo@gmail.com

10. References

[1] http://partsregistry.org/wiki/index.php/Part_Types:Measurement_Systems

[2] Haiyao Huang “Design and Characterization of Artificial Transcriptional Terminators”.

[3] Lars Velten, Nao” Units for Promoter Measurement in Mammalian Cells”