Difference between revisions of "Team:Worldshaper-Wuhan/Measurement"

 
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.tw-social-icon a.dribbble:hover span{background-color:#f06eaa}.tw-social-icon a.linkedin:hover span{background-color:#39c}.tw-social-icon a.instagram:hover span{background-color:#9f6f59}.tw-social-icon a.rss:hover span{background-color:#fca600}#footer .tw-social-icon a,#footer .tw-social-icon a:hover span{background:0}#footer .tw-social-icon a{margin-top:-2px}div.jp-jplayer.jp-jplayer-video{background-color:#292a2b}.jp-jplayer-video video{margin-bottom:-6px}div.jp-audio,div.jp-video{color:#fff;font-size:1em;font-family:Verdana,Arial,sans-serif;line-height:1.6}div.jp-audio,div.jp-video{width:100%}div.jp-interface{position:relative;z-index:100;width:100%;background:url(../img/jplayer_bg.png) repeat-x;-webkit-box-shadow:0 1px 1px rgba(0,0,0,0.4);-moz-box-shadow:0 1px 1px rgba(0,0,0,0.4);box-shadow:0 1px 1px rgba(0,0,0,0.4)}div.jp-type-single div.jp-interface{height:30px;overflow:hidden}div.jp-interface ul.jp-controls{margin:0;padding:0;list-style-type:none}div.jp-interface ul.jp-controls 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        .tw-social-icon a.dribbble:hover span{background-color:#f06eaa}.tw-social-icon a.linkedin:hover span{background-color:#39c}.tw-social-icon a.instagram:hover span{background-color:#9f6f59}.tw-social-icon a.rss:hover span{background-color:#fca600}#footer .tw-social-icon a,#footer .tw-social-icon a:hover span{background:0}#footer .tw-social-icon a{margin-top:-2px}div.jp-jplayer.jp-jplayer-video{background-color:#292a2b}.jp-jplayer-video video{margin-bottom:-6px}div.jp-audio,div.jp-video{color:#fff;font-size:1em;font-family:Verdana,Arial,sans-serif;line-height:1.6}div.jp-audio,div.jp-video{width:100%}div.jp-interface{position:relative;z-index:100;width:100%;background:url(../img/jplayer_bg.png) repeat-x;-webkit-box-shadow:0 1px 1px rgba(0,0,0,0.4);-moz-box-shadow:0 1px 1px rgba(0,0,0,0.4);box-shadow:0 1px 1px rgba(0,0,0,0.4)}div.jp-type-single div.jp-interface{height:30px;overflow:hidden}div.jp-interface ul.jp-controls{margin:0;padding:0;list-style-type:none}div.jp-interface ul.jp-controls li{display:inline}div.jp-interface ul.jp-controls a{position:absolute;overflow:hidden;text-indent:-9999px}a.jp-play,a.jp-pause{z-index:1;width:33px;height:30px;outline:0}div.jp-type-single a.jp-play,div.jp-type-single a.jp-pause{top:0;left:0}a.jp-play{background:url("../img/jplayer-black-and-yellow.png") 0 0 no-repeat}a.jp-play:hover{background:url("../img/jplayer-black-and-yellow.png") -33px 0 no-repeat}a.jp-pause{display:none;background:url("../img/jplayer-black-and-yellow.png") 0 -30px no-repeat}a.jp-pause:hover{background:url("../img/jplayer-black-and-yellow.png") -33px -30px no-repeat}div.jp-progress-container{position:absolute;top:0;left:34px;padding:12px 10px;width:225px;height:6px}div.jp-progress{position:absolute;overflow:hidden}div.jp-type-single div.jp-progress{padding:1px;width:223px;height:7px;background:url("../img/jplayer-black-and-yellow.png") 0 -192px repeat-x}.single div.jp-type-single div.jp-progress{width:411px}div.jp-seek-bar{width:0;height:5px;-webkit-border-radius:1px;-moz-border-radius:1px;border-radius:1px;background:url("../img/jplayer-black-and-yellow.png") 0 -163px repeat-x;cursor:pointer}div.jp-play-bar{width:0;height:5px;-webkit-border-radius:1px;-moz-border-radius:1px;border-radius:1px;background:url("../img/jplayer-black-and-yellow.png") 0 -133px repeat-x}div.jp-volume-bar-container{position:absolute;top:0;right:20px;padding:12px 10px;width:40px;height:6px}div.jp-volume-bar{position:relative;overflow:hidden;width:40px;height:9px;background:url("../img/jplayer-black-and-yellow.png") 0 -192px repeat-x;cursor:pointer}div.jp-volume-bar-value{margin:1px;width:0;height:5px;-webkit-border-radius:1px;-moz-border-radius:1px;border-radius:1px;background:url("../img/jplayer-black-and-yellow.png") 0 -133px 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#fbcomments,.fb-comments,.fb-comments iframe[style],.fb-comments span{width:100%!important}.single .entry-tags i{padding-right:10px}.single .entry-tags a{padding:3px 5px;border:1px solid #ccc;margin-right:5px}.single .pagination li{list-style:none}.single .pagination,.single .pagination ul{margin:0;padding:0}.single .pagination a{width:40px;line-height:40px;display:inline-block;text-align:center;background:#2b353e;color:#fff}.single .pagination a:hover{background:#1d2124}a.btn,#footer a.btn{color:#fff}.btn{overflow:hidden;display:inline-block;margin:0 10px 10px 0;padding:14px 20px;background:#1d2124;font-family:'Lato',sans-serif;font-size:13px;font-weight:bold;line-height:1;text-transform:uppercase;*display:inline;*zoom:1;position:relative;-webkit-transition:all .3s;-moz-transition:all .3s;transition:all .3s;-webkit-border-radius:3px;-moz-border-radius:3px;border-radius:3px}.btn i{margin-right:7px;font-size:14px}.btn-border,.btn-border span{transition:all 300ms;-webkit-transition:all 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.owl-buttons{display:none}#scrollUp{position:fixed;right:30px;bottom:30px;display:none;width:45px;height:45px;color:#FFF;text-align:center;background-color:#21b6a8;font-size:16px;line-height:45px;cursor:pointer;border-radius:50%;-webkit-border-radius:50%;-moz-border-radius:50%}#scrollUp:hover{background-color:#25a196}.background-video,.video-mask,.video-mask-color{position:absolute;width:100%;height:100%}.video-mask{top:0;left:0;z-index:3;cursor:pointer;background-image:url(../img/video-mask.png);background-repeat:repeat;background-size:2px 2px;transition:background 1s;-webkit-transition:background 1s;-moz-transition:background 1s;-o-transition:background 1s}.paused>.video-mask{background-color:rgba(0,0,0,0.5)}.video-mask-color{top:0;left:0;z-index:3;background:#000;zoom:1;filter:alpha(opacity=30);opacity:.3}
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        #fbcomments,.fb-comments,.fb-comments iframe[style],.fb-comments span{width:100%!important}.single .entry-tags i{padding-right:10px}.single .entry-tags a{padding:3px 5px;border:1px solid #ccc;margin-right:5px}.single .pagination li{list-style:none}.single .pagination,.single .pagination ul{margin:0;padding:0}.single .pagination a{width:40px;line-height:40px;display:inline-block;text-align:center;background:#2b353e;color:#fff}.single .pagination a:hover{background:#1d2124}a.btn,#footer a.btn{color:#fff}.btn{overflow:hidden;display:inline-block;margin:0 10px 10px 0;padding:14px 20px;background:#1d2124;font-family:'Lato',sans-serif;font-size:13px;font-weight:bold;line-height:1;text-transform:uppercase;*display:inline;*zoom:1;position:relative;-webkit-transition:all .3s;-moz-transition:all .3s;transition:all .3s;-webkit-border-radius:3px;-moz-border-radius:3px;border-radius:3px}.btn i{margin-right:7px;font-size:14px}.btn-border,.btn-border span{transition:all 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.owl-buttons{display:none}#scrollUp{position:fixed;right:30px;bottom:30px;display:none;width:45px;height:45px;color:#FFF;text-align:center;background-color:#21b6a8;font-size:16px;line-height:45px;cursor:pointer;border-radius:50%;-webkit-border-radius:50%;-moz-border-radius:50%}#scrollUp:hover{background-color:#25a196}.background-video,.video-mask,.video-mask-color{position:absolute;width:100%;height:100%}.video-mask{top:0;left:0;z-index:3;cursor:pointer;background-image:url(../img/video-mask.png);background-repeat:repeat;background-size:2px 2px;transition:background 1s;-webkit-transition:background 1s;-moz-transition:background 1s;-o-transition:background 1s}.paused>.video-mask{background-color:rgba(0,0,0,0.5)}.video-mask-color{top:0;left:0;z-index:3;background:#000;zoom:1;filter:alpha(opacity=30);opacity:.3}
.bg-video-container{text-align:center;margin-top:44px;margin-bottom:46px}.bg-video-play{cursor:pointer;color:#808080;width:150px;height:150px;-webkit-border-radius:50%;-moz-border-radius:50%;border-radius:50%;background-color:#fff;border-style:solid;border-width:12px;border-color:rgba(0,0,0,.33);font-size:40px;line-height:150px;margin:55px auto;position:relative;z-index:1;-webkit-transition:all .4s ease;-moz-transition:all .4s ease;-o-transition:all .4s ease;-ms-transition:all .4s ease;transition:all .4s ease}.bg-video-play:hover{color:#fff;background-color:rgba(0,0,0,.33);border-color:#fff}.bg-video-container h2{color:#fff;font-weight:900;text-transform:uppercase;font-size:48px;line-height:65px}.bg-video-horizontal{position:relative}.bg-video-horizontal h2{position:absolute;width:50%;padding-right:125px;top:50%;margin-top:-70px;font-size:40px}.bg-video-horizontal h2.bg-video-last-text{right:0;padding-right:0;padding-left:125px}#main{overflow:hidden;padding:30px 0}.waves-pagebuilder #main{padding:0}#main,#slider{position:relative;background:#FFF}.page-template-template-onepage-php #main{padding:0}#main [class*="col-md"]{display:inline-block;margin-top:30px;margin-bottom:30px;*display:inline}#main .waves-portfolio [class*="col-md"]{margin-top:15px;margin-bottom:15px}#main .waves-portfolio [class*="col-md-3"]{width:24.9%}.onepage-header-container [class*="col-md"]{display:inline-block;margin-top:30px;margin-bottom:30px;*display:inline}.bg-scroll,.bg-fixed,.bg-parallax{background-size:cover}.bg-fixed,.bg-parallax{background-attachment:fixed}.bg-pattern{background-repeat:repeat;background-position:left top}.waves-about{background-image:url(../img/about-bg.png);background-repeat:repeat-y;background-position:50%}.waves-about .about-type-content{position:relative}.waves-about .about-type-content>.about-bullet{position:absolute;top:50%;left:50%;border:solid 2px #d5d5d5;width:30px;height:30px;border-radius:50%;-webkit-border-radius:50%;-moz-border-radius:50%;margin-top:-15px;margin-left:-15px;background-color:#fff}.waves-about .about-type-content>.about-bullet,.waves-about .about-type-content>.about-content,.waves-about .about-type-content>.about-content:before{-webkit-transition:all .4s ease;-moz-transition:all .4s ease;-o-transition:all .4s ease;-ms-transition:all .4s ease;transition:all .4s ease}.waves-about .about-type-content:hover>.about-bullet{border-color:#fff;background-color:#21b6a8}.waves-about .about-type-title{text-align:center}.waves-about .about-type-title>.about-title{color:#FFF;background-color:#21b6a8;display:inline-block;padding:9px 24px;font-size:20px;line-height:1;-webkit-border-radius:3px;-moz-border-radius:3px;border-radius:3px;border:solid 2px #fff;margin:60px auto}.waves-about .about-type-title:first-child>.about-title{margin-top:0}.waves-about .about-type-title:last-child>.about-title{margin-bottom:0}.waves-about .about-type-content>.about-content,.waves-about .about-type-content>.about-content:before{width:45%;background-color:#f5f5f5;border-style:solid;border-width:1px;border-color:#f5f5f5}.waves-about .about-type-content>.about-content:before{position:absolute;content:"";top:50%;right:-10px;margin-top:-10px;width:20px;height:20px;-webkit-transform:rotate(45deg);-moz-transform:rotate(45deg);-o-transform:rotate(45deg);-ms-transform:rotate(45deg);transform:rotate(45deg);border-left-width:0;border-bottom-width:0}.waves-about .about-pos-right>.about-content:before{right:auto;left:-10px;border-left-width:1px;border-bottom-width:1px;border-right-width:0;border-top-width:0}.waves-about .about-type-content>.about-content{position:relative;color:#808080;padding:16px 28px;border-radius:3px;-webkit-border-radius:3px;-moz-border-radius:3px}.waves-about .about-pos-right>.about-content{float:right}.waves-about .about-type-content .about-heading>div{display:inline-block}.waves-about .about-type-content .about-title{font-family:'Lato';font-size:20px;color:#808080;font-weight:bold}.waves-about .about-type-content .about-date{float:right;font-size:16px}.waves-about .about-type-content:hover .about-content,.waves-about .about-type-content:hover .about-content:before{background-color:#fff;border-color:#d5d5d5}.waves-about .about-type-content:hover .about-title{color:#21b6a8;font-weight:600}.waves-about .about-type-content:hover .about-date{font-weight:600}.waves-aboutme .aboutme-meta .aboutme-share{text-align:right}.waves-aboutme .aboutme-meta .desc{margin-bottom:10px}.waves-aboutme .aboutme-meta>div{width:50%;display:inline-block}.waves-aboutme .aboutme-head{font-size:30px}.waves-aboutme .aboutme-head h2{font-size:43px;margin:10px 0;color:#1c1c1c}.waves-aboutme .aboutme-meta a i{background-color:transparent;border:solid 1px #808080;margin-right:2px;height:43px;width:43px;font-size:20px;text-align:center;vertical-align:middle;padding-top:11px}
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        .bg-video-container{text-align:center;margin-top:44px;margin-bottom:46px}.bg-video-play{cursor:pointer;color:#808080;width:150px;height:150px;-webkit-border-radius:50%;-moz-border-radius:50%;border-radius:50%;background-color:#fff;border-style:solid;border-width:12px;border-color:rgba(0,0,0,.33);font-size:40px;line-height:150px;margin:55px auto;position:relative;z-index:1;-webkit-transition:all .4s ease;-moz-transition:all .4s ease;-o-transition:all .4s ease;-ms-transition:all .4s ease;transition:all .4s ease}.bg-video-play:hover{color:#fff;background-color:rgba(0,0,0,.33);border-color:#fff}.bg-video-container h2{color:#fff;font-weight:900;text-transform:uppercase;font-size:48px;line-height:65px}.bg-video-horizontal{position:relative}.bg-video-horizontal h2{position:absolute;width:50%;padding-right:125px;top:50%;margin-top:-70px;font-size:40px}.bg-video-horizontal h2.bg-video-last-text{right:0;padding-right:0;padding-left:125px}#main{overflow:hidden;padding:30px 0}.waves-pagebuilder #main{padding:0}#main,#slider{position:relative;background:#FFF}.page-template-template-onepage-php #main{padding:0}#main [class*="col-md"]{display:inline-block;margin-top:30px;margin-bottom:30px;*display:inline}#main .waves-portfolio [class*="col-md"]{margin-top:15px;margin-bottom:15px}#main .waves-portfolio [class*="col-md-3"]{width:24.9%}.onepage-header-container [class*="col-md"]{display:inline-block;margin-top:30px;margin-bottom:30px;*display:inline}.bg-scroll,.bg-fixed,.bg-parallax{background-size:cover}.bg-fixed,.bg-parallax{background-attachment:fixed}.bg-pattern{background-repeat:repeat;background-position:left top}.waves-about{background-image:url(../img/about-bg.png);background-repeat:repeat-y;background-position:50%}.waves-about .about-type-content{position:relative}.waves-about .about-type-content>.about-bullet{position:absolute;top:50%;left:50%;border:solid 2px #d5d5d5;width:30px;height:30px;border-radius:50%;-webkit-border-radius:50%;-moz-border-radius:50%;margin-top:-15px;margin-left:-15px;background-color:#fff}.waves-about .about-type-content>.about-bullet,.waves-about .about-type-content>.about-content,.waves-about .about-type-content>.about-content:before{-webkit-transition:all .4s ease;-moz-transition:all .4s ease;-o-transition:all .4s ease;-ms-transition:all .4s ease;transition:all .4s ease}.waves-about .about-type-content:hover>.about-bullet{border-color:#fff;background-color:#21b6a8}.waves-about .about-type-title{text-align:center}.waves-about .about-type-title>.about-title{color:#FFF;background-color:#21b6a8;display:inline-block;padding:9px 24px;font-size:20px;line-height:1;-webkit-border-radius:3px;-moz-border-radius:3px;border-radius:3px;border:solid 2px #fff;margin:60px auto}.waves-about .about-type-title:first-child>.about-title{margin-top:0}.waves-about .about-type-title:last-child>.about-title{margin-bottom:0}.waves-about .about-type-content>.about-content,.waves-about .about-type-content>.about-content:before{width:45%;background-color:#f5f5f5;border-style:solid;border-width:1px;border-color:#f5f5f5}.waves-about .about-type-content>.about-content:before{position:absolute;content:"";top:50%;right:-10px;margin-top:-10px;width:20px;height:20px;-webkit-transform:rotate(45deg);-moz-transform:rotate(45deg);-o-transform:rotate(45deg);-ms-transform:rotate(45deg);transform:rotate(45deg);border-left-width:0;border-bottom-width:0}.waves-about .about-pos-right>.about-content:before{right:auto;left:-10px;border-left-width:1px;border-bottom-width:1px;border-right-width:0;border-top-width:0}.waves-about .about-type-content>.about-content{position:relative;color:#808080;padding:16px 28px;border-radius:3px;-webkit-border-radius:3px;-moz-border-radius:3px}.waves-about .about-pos-right>.about-content{float:right}.waves-about .about-type-content .about-heading>div{display:inline-block}.waves-about .about-type-content .about-title{font-family:'Lato';font-size:20px;color:#808080;font-weight:bold}.waves-about .about-type-content .about-date{float:right;font-size:16px}.waves-about .about-type-content:hover .about-content,.waves-about .about-type-content:hover .about-content:before{background-color:#fff;border-color:#d5d5d5}.waves-about .about-type-content:hover .about-title{color:#21b6a8;font-weight:600}.waves-about .about-type-content:hover .about-date{font-weight:600}.waves-aboutme .aboutme-meta .aboutme-share{text-align:right}.waves-aboutme .aboutme-meta .desc{margin-bottom:10px}.waves-aboutme .aboutme-meta>div{width:50%;display:inline-block}.waves-aboutme .aboutme-head{font-size:30px}.waves-aboutme .aboutme-head h2{font-size:43px;margin:10px 0;color:#1c1c1c}.waves-aboutme .aboutme-meta a i{background-color:transparent;border:solid 1px #808080;margin-right:2px;height:43px;width:43px;font-size:20px;text-align:center;vertical-align:middle;padding-top:11px}
.waves-aboutme .aboutme-meta a i:hover{background-color:#fff;color:#21b6a8;border:solid 1px #21b6a8}.waves-aboutme .aboutme-meta a:first-child i{-webkit-border-radius:3px 0 0 3px;-moz-border-radius:3px 0 0 3px;border-radius:3px 0 0 3px}.waves-aboutme .aboutme-meta a:last-child i{-webkit-border-radius:0 3px 3px 0;-moz-border-radius:0 3px 3px 0;border-radius:0 3px 3px 0}.waves-aboutme.aboutme-style-2 .aboutme-head{margin-bottom:30px}.waves-aboutme.aboutme-style-2 span,.waves-aboutme.aboutme-style-2 h2{font-size:35px;font-weight:300}.waves-aboutme.aboutme-style-2 h2{display:inline-block;margin-left:15px;color:#21b6a8}.waves-aboutme.aboutme-style-2 .aboutme-meta{border-bottom:1px solid #e5e5e5;border-top:1px solid #e5e5e5;padding:10px 0 30px 0;margin-top:60px}.waves-aboutme.aboutme-style-2 span.style2{display:block;font-size:29px}.waves-aboutme.aboutme-style-2 .aboutme-meta a i{background-color:#21b6a8;border:solid 1px #21b6a8;color:#fff}.waves-aboutme.aboutme-style-2 .aboutme-meta a i:hover{background-color:#fff;color:#21b6a8;border:solid 1px #21b6a8}.accordion-group{border-style:solid;border-width:2px;border-color:#e5e5e5;border-top-width:0;border-bottom-width:0}.accordion-group .accordion-heading{border-bottom-style:solid;border-bottom-width:2px;border-bottom-color:#fff}.accordion-group:first-child{border-top-width:2px;-webkit-border-radius:3px 3px 0 0;-moz-border-radius:3px 3px 0 0;border-radius:3px 3px 0 0;overflow:hidden}.accordion-group:last-child{border-bottom-color:#e5e5e5;-webkit-border-radius:0 0 3px 3px;-moz-border-radius:0 0 3px 3px;border-radius:0 0 3px 3px;overflow:hidden;border-bottom-width:2px!important}.accordion-group:last-child .accordion-heading{border-bottom-width:0}.accordion-heading .accordion-toggle{position:relative;display:block;padding:20px 23px 15px;cursor:pointer;font-family:'Lato';line-height:1;font-weight:300;font-size:20px;color:#808080;background-color:#f5f5f5;-webkit-transition:none;-moz-transition:none;-o-transition:none;-ms-transition:none;transition:none}.accordion-heading .accordion-toggle:hover{color:#1d2124}.accordion-inner{padding:17px 22px}.accordion-inner p:last-child{margin-bottom:0}.collapse{position:relative;display:none;overflow:hidden}.accordion-group.active .collapse,.accordion-group.closing .collapse{display:block}.accordion-group.active,.accordion-group.active .accordion-heading{border-width:0}.accordion-group.active .accordion-body{border-color:#e5e5e5;border-left-width:2px;border-left-style:solid;border-right-width:2px;border-right-style:solid}.accordion-group.active .accordion-toggle{background-color:#21b6a8;color:#fff;font-weight:700}.waves-callout .callout-container{-webkit-border-radius:5px;-moz-border-radius:5px;border-radius:5px;background-color:#21b6a8;-webkit-box-shadow:4px 4px #1a9d91;-moz-box-shadow:4px 4px #1a9d91;box-shadow:4px 4px #1a9d91;text-align:center;padding:35px 60px;margin:0 30px}.waves-callout .callout-text{position:relative}.waves-callout .callout-text p{font-size:24px;line-height:1.4;margin-bottom:0;color:#FFF}.waves-callout .callout-text p b{font-weight:900}.waves-callout.with-button p{padding-right:180px}.waves-callout .btn-callout{position:absolute;top:50%;right:0;margin:-24px 0 0 20px;padding:12px 30px;font-size:20px;text-transform:uppercase;font-weight:900;border:2px solid transparent;background:rgba(0,0,0,.3);color:#fff}.waves-callout .callout-text>a.btn:hover{background:0;border-color:#fff}.waves-callout .callout-text>a.btn i{margin:0 0 0 10px}.waves-callout.style2 .callout-container{-webkit-border-radius:0;-moz-border-radius:0;border-radius:0;-webkit-box-shadow:none;-moz-box-shadow:none;box-shadow:none;text-align:left;padding:35px 50px}.waves-callout.style2 .callout-text p b{font-weight:700}.waves-callout.style2 .btn-callout{background:#fff;color:#21b6a8;font-size:18px;font-weight:bold;padding:16px 30px;-webkit-box-shadow:3px 3px #1a9d91;-moz-box-shadow:3px 3px #1a9d91;box-shadow:3px 3px #1a9d91}.waves-callout.style2 .btn-callout:hover{color:#fff}.owl-carousel .owl-wrapper:after{content:".";display:block;clear:both;visibility:hidden;line-height:0;height:0}.list_carousel .owl-buttons div{display:inline-block;padding:0 5px;font-size:16px}.owl-theme .owl-controls .owl-page{display:inline-block;zoom:1;*display:inline}.owl-theme .owl-controls .owl-page span{display:block;width:12px;height:12px;margin:0 3px;background:url(../img/owl-page.png);background-position:12px 0}.owl-theme .owl-controls .owl-page.active span,.owl-theme .owl-controls.clickable .owl-page:hover span{background-position:12px 12px}.waves-carousel-twitter.list_carousel .owl-buttons,.waves-carousel-testimonial.list_carousel 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        .waves-aboutme .aboutme-meta a i:hover{background-color:#fff;color:#21b6a8;border:solid 1px #21b6a8}.waves-aboutme .aboutme-meta a:first-child i{-webkit-border-radius:3px 0 0 3px;-moz-border-radius:3px 0 0 3px;border-radius:3px 0 0 3px}.waves-aboutme .aboutme-meta a:last-child i{-webkit-border-radius:0 3px 3px 0;-moz-border-radius:0 3px 3px 0;border-radius:0 3px 3px 0}.waves-aboutme.aboutme-style-2 .aboutme-head{margin-bottom:30px}.waves-aboutme.aboutme-style-2 span,.waves-aboutme.aboutme-style-2 h2{font-size:35px;font-weight:300}.waves-aboutme.aboutme-style-2 h2{display:inline-block;margin-left:15px;color:#21b6a8}.waves-aboutme.aboutme-style-2 .aboutme-meta{border-bottom:1px solid #e5e5e5;border-top:1px solid #e5e5e5;padding:10px 0 30px 0;margin-top:60px}.waves-aboutme.aboutme-style-2 span.style2{display:block;font-size:29px}.waves-aboutme.aboutme-style-2 .aboutme-meta a i{background-color:#21b6a8;border:solid 1px #21b6a8;color:#fff}.waves-aboutme.aboutme-style-2 .aboutme-meta a i:hover{background-color:#fff;color:#21b6a8;border:solid 1px #21b6a8}.accordion-group{border-style:solid;border-width:2px;border-color:#e5e5e5;border-top-width:0;border-bottom-width:0}.accordion-group .accordion-heading{border-bottom-style:solid;border-bottom-width:2px;border-bottom-color:#fff}.accordion-group:first-child{border-top-width:2px;-webkit-border-radius:3px 3px 0 0;-moz-border-radius:3px 3px 0 0;border-radius:3px 3px 0 0;overflow:hidden}.accordion-group:last-child{border-bottom-color:#e5e5e5;-webkit-border-radius:0 0 3px 3px;-moz-border-radius:0 0 3px 3px;border-radius:0 0 3px 3px;overflow:hidden;border-bottom-width:2px!important}.accordion-group:last-child .accordion-heading{border-bottom-width:0}.accordion-heading .accordion-toggle{position:relative;display:block;padding:20px 23px 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#1a9d91;-moz-box-shadow:4px 4px #1a9d91;box-shadow:4px 4px #1a9d91;text-align:center;padding:35px 60px;margin:0 30px}.waves-callout .callout-text{position:relative}.waves-callout .callout-text p{font-size:24px;line-height:1.4;margin-bottom:0;color:#FFF}.waves-callout .callout-text p b{font-weight:900}.waves-callout.with-button p{padding-right:180px}.waves-callout .btn-callout{position:absolute;top:50%;right:0;margin:-24px 0 0 20px;padding:12px 30px;font-size:20px;text-transform:uppercase;font-weight:900;border:2px solid transparent;background:rgba(0,0,0,.3);color:#fff}.waves-callout .callout-text>a.btn:hover{background:0;border-color:#fff}.waves-callout .callout-text>a.btn i{margin:0 0 0 10px}.waves-callout.style2 .callout-container{-webkit-border-radius:0;-moz-border-radius:0;border-radius:0;-webkit-box-shadow:none;-moz-box-shadow:none;box-shadow:none;text-align:left;padding:35px 50px}.waves-callout.style2 .callout-text p b{font-weight:700}.waves-callout.style2 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+
        .process-number{position:absolute;top:0;left:35px;font-weight:normal;font-size:16px;width:40px;height:40px;line-height:40px;background:#2b353e;color:#fff}.process-title{margin:0 auto;width:120px;height:120px;line-height:1;font-size:14px;padding:50px 20px}@-webkit-keyframes progress-bar-stripes{from{background-position:40px 0}to{background-position:0 0}}@-moz-keyframes progress-bar-stripes{from{background-position:40px 0}to{background-position:0 0}}@-ms-keyframes progress-bar-stripes{from{background-position:40px 0}to{background-position:0 0}}@-o-keyframes progress-bar-stripes{from{background-position:0 0}to{background-position:40px 0}}@keyframes progress-bar-stripes{from{background-position:40px 0}to{background-position:0 0}}.progress{overflow:hidden;height:16px;margin-bottom:16px;background-color:#f7f7f7;background-image:-moz-linear-gradient(top,#f5f5f5,#f9f9f9);background-image:-webkit-gradient(linear,0 0,0 100%,from(#f5f5f5),to(#f9f9f9));background-image:-webkit-linear-gradient(top,#f5f5f5,#f9f9f9);background-image:-o-linear-gradient(top,#f5f5f5,#f9f9f9);background-image:linear-gradient(to bottom,#f5f5f5,#f9f9f9);background-repeat:repeat-x;filter:progid:DXImageTransform.Microsoft.gradient(startColorstr='#fff5f5f5',endColorstr='#fff9f9f9',GradientType=0);-webkit-box-shadow:inset 0 1px 2px rgba(0,0,0,0.1);-moz-box-shadow:inset 0 1px 2px rgba(0,0,0,0.1);box-shadow:inset 0 1px 2px rgba(0,0,0,0.1);-webkit-border-radius:4px;-moz-border-radius:4px;border-radius:4px}.waves-progress .bar{width:0;height:12px;color:#fff;font-size:12px;text-align:center;text-shadow:0 -1px 0 rgba(0,0,0,0.25);background-color:#0e90d2;background-repeat:repeat-x;-webkit-transition:width .6s ease;-moz-transition:width .6s ease;-o-transition:width .6s ease;transition:width .6s ease}.waves-progress .bar+.bar{-webkit-box-shadow:inset 1px 0 0 rgba(0,0,0,.15),inset 0 -1px 0 rgba(0,0,0,.15);-moz-box-shadow:inset 1px 0 0 rgba(0,0,0,.15),inset 0 -1px 0 rgba(0,0,0,.15);box-shadow:inset 1px 0 0 rgba(0,0,0,.15),inset 0 -1px 0 rgba(0,0,0,.15)}.progress-striped .bar{background-color:#149bdf;background-image:-webkit-gradient(linear,0 100%,100% 0,color-stop(0.25,rgba(255,255,255,0.15)),color-stop(0.25,transparent),color-stop(0.5,transparent),color-stop(0.5,rgba(255,255,255,0.15)),color-stop(0.75,rgba(255,255,255,0.15)),color-stop(0.75,transparent),to(transparent));background-image:-webkit-linear-gradient(45deg,rgba(255,255,255,0.15) 25%,transparent 25%,transparent 50%,rgba(255,255,255,0.15) 50%,rgba(255,255,255,0.15) 75%,transparent 75%,transparent);background-image:-moz-linear-gradient(45deg,rgba(255,255,255,0.15) 25%,transparent 25%,transparent 50%,rgba(255,255,255,0.15) 50%,rgba(255,255,255,0.15) 75%,transparent 75%,transparent);background-image:-o-linear-gradient(45deg,rgba(255,255,255,0.15) 25%,transparent 25%,transparent 50%,rgba(255,255,255,0.15) 50%,rgba(255,255,255,0.15) 75%,transparent 75%,transparent);background-image:linear-gradient(45deg,rgba(255,255,255,0.15) 25%,transparent 25%,transparent 50%,rgba(255,255,255,0.15) 50%,rgba(255,255,255,0.15) 75%,transparent 75%,transparent);-webkit-background-size:40px 40px;-moz-background-size:40px 40px;-o-background-size:40px 40px;background-size:40px 40px}.waves-progress.active .bar{-webkit-animation:progress-bar-stripes 2s linear infinite;-moz-animation:progress-bar-stripes 2s linear infinite;-ms-animation:progress-bar-stripes 2s linear infinite;-o-animation:progress-bar-stripes 2s linear infinite;animation:progress-bar-stripes 2s linear infinite}.waves-progress .bar{-webkit-border-radius:3px;-moz-border-radius:3px;border-radius:3px}.waves-progress .bar-container{overflow:hidden;background-color:#f5f5f5;-webkit-border-radius:3px;-moz-border-radius:3px;border-radius:3px}.waves-progress{margin-bottom:35px}.waves-progress:last-child{margin-bottom:0}.progress-title{font-weight:300;position:relative;font-size:16px;line-height:1;margin-bottom:13px}.progress-title span{font-weight:700;float:right}.progress .bar.tw-bi{background-image:none}.tw-service-box{padding-bottom:30px}.tw-service-box:last-child{padding-bottom:0}.tw-service-box .tw-service-icon{display:block;margin-bottom:28px;text-align:center}.tw-service-box .tw-service-icon.pull-left,.tw-service-box .tw-service-icon.pull-right{margin-bottom:10px}.tw-service-box .tw-service-content.desc_unstyle{text-align:inherit}.tw-service-box .tw-service-content{text-align:center}.tw-service-box .tw-service-content p:last-child{margin-bottom:0}.tw-service-box .tw-service-content a:hover{color:#222}.tw-service-box h3{cursor:pointer;margin-bottom:15px;font-size:20px;font-weight:900;line-height:1;color:#808080}.tw-service-content a.more{-webkit-border-radius:3px;-moz-border-radius:3px;border-radius:3px;border:solid 2px #808080;font-family:'Lato';font-size:15px;font-weight:bold;color:#808080;padding:9px 20px;text-transform:uppercase;margin-top:25px;display:inline-block}.tw-service-content a.more:hover{border-color:#21b6a8;background-color:#21b6a8;color:#fff}
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a.more{border-color:#21b6a8;background-color:#21b6a8;color:#fff}.tw-service-box.style_1.left-service:hover .tw-service-content a.more{color:#21b6a8;background-color:transparent!important}.tw-service-box.style_2:hover h3,.tw-service-box.style_1:hover h3{color:#21b6a8}.tw-service-box.style_1:hover .tw-font-icon{color:#fff!important;background-color:#21b6a8!important;border-color:#21b6a8!important}.tw-service-box.style_2:hover .tw-font-icon{color:#fff!important;background-color:#21b6a8!important;border-color:#fff!important}.tw-service-box.style_2:hover{background-color:#f5f5f5;border:solid 1px 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        div.wpcf7{position:relative}.wpcf7 input[type="text"],.wpcf7 input[type="password"],.wpcf7 input[type="email"],.wpcf7 textarea{margin:10px 0;padding:10px 15px;-webkit-border-radius:2px;-moz-border-radius:2px;border-radius:2px;background:#f5f5f5}.wpcf7 input[type="text"],.wpcf7 input[type="password"],.wpcf7 input[type="email"]{display:inline-block;position:relative;width:49.7%}.wpcf7 textarea{max-height:163px;position:absolute;right:0;top:-184px;width:48%;min-width:500px}span.wpcf7-form-control-wrap{position:relative;display:block}.wpcf7 input[type="submit"],.wpcf7 input[type="submit"]:hover{margin:0 auto;padding:15px 30px 15px 50px;-webkit-border-radius:2px;-moz-border-radius:2px;border-radius:2px;font-weight:bold;text-transform:uppercase;border:2px solid #808080;color:#808080}.wpcf7 input[type="submit"]:hover{opacity:.9}.wpcf7 .wpcf7-submit{background:url(../img/submit.png) 20px 12px no-repeat;display:block}span.wpcf7-not-valid-tip{color:#f35050;font-size:16px;font-weight:500;text-transform:uppercase;display:inline-block;position:absolute;top:5px;left:15px;background:#f5f5f5}div.wpcf7-validation-errors{border:2px solid #f7e700;margin:0 auto;width:60%;font-weight:bold;text-align:center}.tw-element.waves-shortcode{display:inline-block;*display:inline}#sidebar aside.widget ul.menu li{display:block}aside.widget ul.menu li[class*="fa-"] a{padding-left:10px}aside.widget ul.menu li:before{font-family:'Fontawesome';width:18px;display:inline-block;font-size:14px;text-align:center}body.menu-right.theme-boxed{margin-right:300px}body.menu-left.theme-boxed{margin-left:300px}#waves-sidebar{position:fixed;overflow:hidden;top:0;left:0;height:100%;width:300px}body.admin-bar #waves-sidebar{top:32px}#waves-sidebar.right-sidebar{left:auto;right:0}.sidebar-content{padding:30px;background:#efefef}#waves-sidebar:hover>.pane,.pane.active,.pane.flashed{visibility:visible\9;opacity:.99;filter:alpha(opacity=99);zoom:1}#waves-sidebar>.pane{position:absolute;top:5px;right:5px;bottom:5px;width:10px;visibility:hidden\9;opacity:.01;filter:alpha(opacity=1);zoom:1;background-color:black;background-color:rgba(0,0,0,0.075);border-radius:5px;-webkit-transition:opacity .3s ease;transition:opacity .3s ease}#waves-sidebar.left-sidebar>.pane{right:auto;left:5px}#waves-sidebar>.pane>.slider{position:relative;background-color:rgba(0,0,0,0.1)}#waves-sidebar .sidebar-content{background:#fff;overflow:scroll;overflow-x:hidden;position:absolute;top:0;left:0;right:0;bottom:0;border-right:1px solid #EEE}#left{float:left;width:300px}.waves-container{position:relative}.waves-container>.waves-main,.waves-container>.row>.waves-main{padding:30px 15px}.waves-container>.waves-sidebar.left-sidebar,.waves-container>.row>.waves-sidebar.left-sidebar{padding-top:30px;padding-left:15px}.waves-container>.waves-sidebar.right-sidebar,.waves-container>.row>.waves-sidebar.right-sidebar{padding-top:30px;padding-right:15px}.waves-main.left{margin-right:300px}.waves-main.right{margin-left:300px}.waves-sidebar{padding-left:15px;position:absolute;top:0;right:0;width:300px}.waves-sidebar.left-sidebar{padding-left:0;padding-right:15px;right:auto;left:0}input[type="text"].wysija-input{padding:0 10px;margin-top:10px}.widget_wysija_cont .wysija-submit{border-color:#999;color:#bbb;padding:10px}body.loading{overflow:hidden}body.pace-done>.pace,.pace-inactive,body.pace-done #waves-loader{display:none}.pace .pace-progress{height:4px;background-color:#21b6a8}.theme-boxed{background-image:url(../img/patterns/9.png);background-attachment:fixed;background-repeat:repeat;margin-top:60px;margin-bottom:60px}#div_0e1d_0{background-image:url(../upload/parallax.jpg)}#div_8b27_7{background-image:url(../upload/home-1-parallax.jpg)}#div_0e1d_3{background-image:url(../upload/bg-for-video.jpg);overflow:hidden;position:relative}#div_abc7_20{background-image:url(../upload/bg-parallax-blackpattern.jpg)}#div_cf0e_0{background-image:url(../upload/bg-craft.png)}#p_cf0e_0{text-align:center}#p_cf0e_1{text-align:center}#span_cf0e_0{color:#fff}#p_cf0e_2{text-align:center}#a_cf0e_0{border-color:#10978a;background-color:#10978a}#div_abc7_0{margin-left:47px}#div_abc7_1{margin-left:-47px}#i_abc7_0{border-style:solid;font-style:normal;text-align:center;font-size:30px;width:32px;line-height:32px;padding:29px;color:#808080;background-color:#fff;border-color:#808080;border-width:3px;border-radius:50px;-webkit-border-radius:50px;-moz-border-radius:50px}#div_abc7_2{margin-left:72px}#div_abc7_3{margin-left:47px}#div_abc7_4{margin-left:-47px}#i_abc7_1{border-style:solid;font-style:normal;text-align:center;font-size:30px;width:32px;line-height:32px;padding:29px;color:#808080;background-color:#fff;border-color:#808080;border-width:3px;border-radius:50px;-webkit-border-radius:50px;-moz-border-radius:50px}
#div_abc7_5{margin-left:72px}#div_abc7_6{margin-left:47px}#div_abc7_7{margin-left:-47px}#i_abc7_2{border-style:solid;font-style:normal;text-align:center;font-size:30px;width:32px;line-height:32px;padding:29px;color:#808080;background-color:#fff;border-color:#808080;border-width:3px;border-radius:50px;-webkit-border-radius:50px;-moz-border-radius:50px}#div_abc7_8{margin-left:72px}#div_abc7_9{margin-left:47px}#div_abc7_10{margin-left:-47px}#i_abc7_3{border-style:solid;font-style:normal;text-align:center;font-size:30px;width:32px;line-height:32px;padding:29px;color:#808080;background-color:#fff;border-color:#808080;border-width:3px;border-radius:50px;-webkit-border-radius:50px;-moz-border-radius:50px}#div_abc7_11{margin-left:72px}#div_abc7_12{margin-left:47px}#div_abc7_13{margin-left:-47px}#i_abc7_4{border-style:solid;font-style:normal;text-align:center;font-size:30px;width:32px;line-height:32px;padding:29px;color:#808080;background-color:#fff;border-color:#808080;border-width:3px;border-radius:50px;-webkit-border-radius:50px;-moz-border-radius:50px}#div_abc7_14{margin-left:72px}#div_abc7_15{margin-left:47px}#div_abc7_16{margin-left:-47px}#i_abc7_5{border-style:solid;font-style:normal;text-align:center;font-size:30px;width:32px;line-height:32px;padding:29px;color:#808080;background-color:#fff;border-color:#808080;border-width:3px;border-radius:50px;-webkit-border-radius:50px;-moz-border-radius:50px}#div_abc7_17{margin-left:72px}#div_abc7_18{display:block;margin-bottom:40px;margin-top:60px}#div_abc7_19{background-color:#dbdbdb}#a_abc7_0{border-color:#fff;color:#fff}#span_abc7_0{background-color:#fff}#div_abc7_21{display:block;margin-bottom:40px;margin-top:40px}#div_abc7_22{opacity:0}#div_abc7_23{display:block;margin-bottom:40px;margin-top:40px}#div_abc7_24{margin-left:60px}#div_abc7_25{margin-left:-60px;margin-top:-60px}#i_abc7_6{border-style:solid;font-style:normal;text-align:center;font-size:40px;width:42px;line-height:42px;padding:37px;color:#808080;background-color:#fff;border-color:#808080;border-width:3px;border-radius:74px;-webkit-border-radius:74px;-moz-border-radius:74px}#div_abc7_26{margin-left:85px}#div_abc7_27{margin-left:60px}#div_abc7_28{margin-left:-60px;margin-top:-60px}#i_abc7_7{border-style:solid;font-style:normal;text-align:center;font-size:40px;width:42px;line-height:42px;padding:37px;color:#808080;background-color:#fff;border-color:#808080;border-width:3px;border-radius:74px;-webkit-border-radius:74px;-moz-border-radius:74px}#div_abc7_29{margin-left:85px}#div_abc7_30{margin-left:60px}#div_abc7_31{margin-left:-60px;margin-top:-60px}#i_abc7_8{border-style:solid;font-style:normal;text-align:center;font-size:40px;width:42px;line-height:42px;padding:37px;color:#808080;background-color:#fff;border-color:#808080;border-width:3px;border-radius:74px;-webkit-border-radius:74px;-moz-border-radius:74px}#div_abc7_32{margin-left:85px}#div_abc7_33{display:block;margin-bottom:0;margin-top:30px}#layerslider_6_2{width:1280px;height:720px;max-width:1280px;margin:0 auto;margin-bottom:0}#img_7fb0_0{top:142px;left:566px;white-space:nowrap}#img_7fb0_1{top:350px;left:236px;font-weight:300;font-family:Lato,'Open Sans',sans-serif;font-size:40px;color:#286e89;white-space:nowrap}#img_7fb0_2{top:371px;left:523px;font-weight:500;font-family:Lato,'Open Sans',sans-serif;font-size:30px;color:#fa6b4b;white-space:nowrap}#div_7fb0_0{display:block;margin-bottom:30px;margin-top:0}#div_7fb0_1{background-color:#dbdbdb}#div_7fb0_2{margin-top:68px}#div_7fb0_3{margin-top:-68px}#i_7fb0_0{border-style:solid;font-style:normal;text-align:center;font-size:28px;width:30px;line-height:30px;padding:51px;color:#808080;background-color:fff;border-color:#808080;border-width:3px;border-radius:100px;-webkit-border-radius:100px;-moz-border-radius:100px}#div_7fb0_4{margin-top:68px}#div_7fb0_5{margin-top:-68px}#i_7fb0_1{border-style:solid;font-style:normal;text-align:center;font-size:28px;width:30px;line-height:30px;padding:51px;color:#808080;background-color:#fff;border-color:#808080;border-width:3px;border-radius:100px;-webkit-border-radius:100px;-moz-border-radius:100px}#div_7fb0_6{margin-top:68px}#div_7fb0_7{margin-top:-68px}#i_7fb0_2{border-style:solid;font-style:normal;text-align:center;font-size:28px;width:30px;line-height:30px;padding:51px;color:#808080;background-color:#fff;border-color:#808080;border-width:3px;border-radius:100px;-webkit-border-radius:100px;-moz-border-radius:100px}#div_7fb0_8{margin-top:68px}#div_7fb0_9{margin-top:-68px}#i_7fb0_3{border-style:solid;font-style:normal;text-align:center;font-size:28px;width:30px;line-height:30px;padding:51px;color:#808080;background-color:#fff;border-color:#808080;border-width:3px;border-radius:100px;-webkit-border-radius:100px;-moz-border-radius:100px}#a_0e1d_0{border-color:#21b6a8;color:#21b6a8}#span_0e1d_0{background-color:#21b6a8}#div_0e1d_1{display:block;margin-bottom:80px;margin-top:80px}#div_0e1d_2{margin-bottom:-50px}#div_0e1d_4{position:relative;z-index:4}
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                                        <h2 class="entry-title"><a href="#">Measurement</a></h2>
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                                            <p><strong><span style="font-weight:bold;font-size:19px">Aim of experiment</span></strong><span style="font-size:16px">&nbsp;</span></p><p style="text-align:left"><span style="font-size:16px">Based on J33201 part, a biosensor pSB1C3-pArsR-RBS-GFP(K3153000) was constructed to detect arsenite(As</span><sup><span style="vertical-align:super">3+</span></sup><span style="font-size:16px">) in water and contributed to this well-characterized part.</span><span style="font-size:16px">&nbsp;</span></p><p style="text-align:left"><strong><span style="font-weight:bold;font-size:19px">Methods</span></strong></p><p style="text-align:left"><span style="font-weight:normal;font-size:16px">1. </span><span style="font-weight:normal;font-size:16px;background:rgb(255,255,255)">Growth curve of </span><em><span style="font-weight:normal;font-style:italic;font-size:16px;background:rgb(255,255,255)">E.</span></em><span style="font-weight:normal;font-size:16px;background:rgb(255,255,255)">coli containing part K577881 in different concentrations of arsenite(</span><strong><span style="font-weight:bold;font-size:12px">As</span></strong><strong><sup><span style="font-weight:bold;vertical-align:super">3+</span></sup></strong><span style="font-weight:normal;font-size:16px;background:rgb(255,255,255)">) </span></p><p style="text-align:left"><span style="font-weight:normal;font-size:16px;background:rgb(255,255,255)">Constructed plasmid pSB1C3-pArsR-GFP containing J33201 part was transformed into E.coli DH5α strain. Single colony was selected to inoculate LB broth containing chloramphenicol and cultured overnight. Then overnight culture was inoculated in the fresh LB medium containing 34 μg/ml chloramphenicol at a ratio of 1:100, mixed well and divided into tubes. Different concentrations of arsenite (As<sup><span style="font-weight:bold;vertical-align:super">3+</span></sup>) solutions were added into the test tubes, respectively. The final concentration of arsenic was (0, 5ppb, 10ppb, 50ppb, 100ppb, 500ppb, 1ppm, 5ppm, 10ppm, 50ppm and 100ppm). Samples were collected at different time points of 0 h, 1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 7 h, 8 h and 16 h. OD600 values were measured by the Multiskan Spectrum Microplate Reader. </span></p><p style="text-align:left"><span style="font-weight:normal;font-size:16px;background:rgb(255,255,255)">2. </span><span style="font-weight:normal;font-size:16px;background:rgb(255,255,255)">GFP expression of E.coli under the control of J33201 induced by different concentration of arsenite (As</span><sup><span style="font-weight:normal;vertical-align:super;background:rgb(255,255,255)">3+</span></sup><span style="font-weight:normal;font-size:16px;background:rgb(255,255,255)">)</span></p><p><span style="font-size:16px">Overnight cultured bacterial solution was inoculated in LB broth containing chloramphenicol at 1:50 to expand the culture. When OD600 reached 0.4-0.6, experiments were performed . Different concentrations of arsenite were added into the test tube, respectively. The final concentration of arsenic was 0, 5 ppb, 10 ppb, 50 ppb, 100 ppb, 500 ppb, 1 ppm, 5 ppm, 10 ppm, 50 ppm, 100 ppm. </span></p><p><span style="font-size:16px">(1) Samples were collected at 16 h. GFP fluorescence intensity (485 nm excitation/ 528 nm emission) and OD600 value were measured at the same time. The bacteria were centrifuged, and the pellets were was observed and photographed under Blue Light Gel Imager.</span></p><p><span style="">(2)Samples were </span><span style="font-size:16px">collected</span><span style="">&nbsp;at different time points of 0 h, 2 h, 4 h, 6 h and 16 h.</span><span style="">&nbsp;GFP fluorescence intensity (485 nm excitation/ 528 nm emission) and OD600 value were measured at the same time.</span></p><p><span style="font-size:12px">&nbsp;</span></p><p><strong><span style="font-weight:bold;font-size:19px">Results</span></strong></p><p><span style="font-size:16px">Fig.1 showed that bacteria growth was not affected from 5 ppb to 10 ppm of arsenite solution (As</span><sup><span style="vertical-align:super">3+</span></sup><span style="font-size:16px">). However, when the concentration of arsenite increased to 50 ppm, the growth of the bacteria was seriously inhibited, suggesting the toxictity to cells at this concentration.</span></p><p style="text-align:center"><img alt="Characterization(图1)" style="max-width:100%!important;height:auto;" src="https://2019.igem.org/wiki/images/9/9c/T--Worldshaper-Wuhan--GC-fig1.jpeg" width="547" height="333"></p><p><span style="font-size:16px">&nbsp;</span><br></p><p><strong><span style="font-weight:bold;font-size:12px">Fig.1 Growth curve of E.coli in different concentration of arsenic (As</span></strong><strong><sup><span style="font-weight:bold;vertical-align:super">3+</span></sup></strong><strong><span style="font-weight:bold;font-size:12px">)</span></strong><span style="font-size:12px">.</span></p><p><span style="font-size:16px">&nbsp;</span></p><p><span style="font-size:16px">As shown in Fig.2, our biosensor based on J33201 part is very sensitive to arsenite(As</span><sup><span style="vertical-align:super">3+</span></sup><span style="font-size:16px">) and the fluoresence signal was be detected at 50 ppb arsenite. Significant dose-dependent effect was observed from 50 ppb to 1 ppm arsenite. The threshold of this biosensor is 10 ppm. As the arsenite concentration increased to 50 ppm, it does’t work because of the toxicity to cell at thisconcentration.</span></p><p style="text-align:center"><img alt="Characterization(图2)" style="max-width:100%!important;height:auto;" src="https://2019.igem.org/wiki/images/b/b5/T--Worldshaper-wuhan--GFP1-fig2.jpeg" width="551" height="369"></p><p><span style="font-size:16px">&nbsp;</span><br></p><p><span style="font-size:16px">(Fig.2a)</span></p><p style="text-align:center"><img alt="Characterization(图3)" style="max-width:100%!important;height:auto;" src="https://2019.igem.org/wiki/images/3/33/T--Worldshaper-wuhan--GFP1-fig3.jpeg" width="361" height="111"></p><p><span style="font-size:16px">&nbsp;</span><br></p><p><span style="font-size:16px">(Fig.2b)</span></p><p><strong><span style="font-weight:bold">Fig.2 GFP expression of E.coli under the control of J33201 in different concentration of arsenite (As<sup><span style="vertical-align:super">3+</span></sup>).</span></strong></p><p><span style="font-size:16px">&nbsp;</span></p><p><span style="font-size:16px">As shown in Fig.3, our biosensor by J33201 reacted rapidly in arsenite solution. After 4 h following arsenite treated, the corresponding fluorescence signal was detected at 100 ppb. As time increases, a weak signal can be detected at 50ppb, indicating that the biosensor can work within 4 hours. </span></p><p style="text-align:center"><img alt="Characterization(图4)" style="max-width:100%!important;height:auto;" src="https://2019.igem.org/wiki/images/3/36/T--Worldshaper-wuhan--GFP2-fig4.jpeg" width="548" height="369"></p><p><span style="font-size:16px">&nbsp;</span><br></p><p><strong><span style="font-weight:bold">Fig.3 GFP expression of E.coli in different concentration of arsenite (As<sup><span style="vertical-align:super">3+</span></sup>) at different time points under the control of J33201.</span></strong></p><p><span style="font-size:16px">&nbsp;</span></p><p><strong><span style="font-weight:bold;font-size:19px">Summary</span></strong></p><p><span style="font-size:16px">our biosensor constructed on the basis of J33201 is sens</span><span style="font-size:16px">itive and fast. The biosensor can detect 100ppb-10ppm arsenite (As<sup><span style="vertical-align:super">3+</span></sup>) in water within 4 hours.</span><span style="font-size:16px">&nbsp;</span><span style="font-size:16px">With the extension of time, the minimum detection limit can reach 50 ppb.</span></p><p><strong><span style="font-weight:bold;font-size:19px">Reference</span></strong></p><p style="margin-top:6px;margin-bottom:8px;line-height:24px;background:rgb(255,255,255)"><span style="color:rgb(40,40,40);font-size:13px">Anal Bioanal Chem. 2011 May;400(4):1031-9. Epub 2011 Mar 27.</span></p><p><br></p>
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<p><span style=";font-family:宋体;font-size:19px;background:rgb(255,255,255)">1. Growth</span><span style=";font-family:Calibri;font-size:19px;background:rgb(255,255,255)">&nbsp;curve of E.coli containing part K577881 in different concentration of arsenite </span><span style=";font-family:宋体;font-size:19px;background:rgb(255,255,255)">(</span><span style=";font-family:Calibri;font-size:19px;background:rgb(255,255,255)">arsenite</span><span style=";font-family:宋体;font-size:19px;background:rgb(255,255,255)">)</span><span style=";font-family:Calibri;font-size:14px">&nbsp;</span></p><p><span style=";font-family:宋体;font-size:16px">Fig.</span><span style=";font-family:Calibri;font-size:16px">1</span><span style=";font-family:宋体;font-size:16px">&nbsp;showed that the growth of bacteria was not affected in the range of </span><span style=";font-family:Calibri;font-size:16px">5ppb-10ppm of arsenite solution (As</span><sup><span style=";font-family:Calibri;font-size:16px;vertical-align:super">3+</span></sup><span style=";font-family:Calibri;font-size:16px">)</span><span style=";font-family:宋体;font-size:16px">. </span><span style=";font-family:Calibri;font-size:16px">However, as the concentration of arsenite increased to 50ppm, the growth of the bacteria was seriously inhibited, suggesting the toxictity to cells at this level.</span></p><p><span style=";font-family:Calibri;font-size:14px">Fig.1 </span></p><p style="text-align:center"><img alt="Characterization(图1)" width="554" height="336" src="https://2019.igem.org/wiki/images/9/9c/T--Worldshaper-Wuhan--GC-fig1.jpeg" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"></p><p><span style=";font-family:Calibri;font-size:14px">&nbsp;</span><br></p><p><strong><span style="font-family: Calibri;font-size: 12px">Fig.1 Growth curve of E.coli in different concentration of arsenic (As</span></strong><strong><sup><span style="font-family: Calibri;font-size: 12px;vertical-align: super">3+</span></sup></strong><strong><span style="font-family: Calibri;font-size: 12px">)</span></strong><span style=";font-family:Calibri;font-size:12px">. </span><span style=";font-family:Calibri;font-size:12px">Constructed plasmid</span><span style=";font-family:Calibri;font-size:12px">&nbsp;pSB1C3-pArsR-GFP containing J33201 part</span><span style=";font-family:Calibri;font-size:12px">&nbsp;was transformed into </span><span style=";font-family:Calibri;font-size:12px">E.coli DH5α</span><span style=";font-family:Calibri;font-size:12px">&nbsp;strain. Single colony was selected to inoculate LB broth containing chloramphenicol and cultured overnight. Then overnight culture was inoculated in the fresh LB medium containing chloramphenicol 34</span><span style=";font-family:Calibri;font-size:12px">μ</span><span style=";font-family:Calibri;font-size:12px">g/ml at a ratio of 1:100, mixed well and divide into tubes. Different concentrations of arsenite(As</span><sup><span style=";font-family:Calibri;font-size:12px;vertical-align:super">3+</span></sup><span style=";font-family:Calibri;font-size:12px">) solutions were added into the test tube</span><span style=";font-family:宋体;font-size:12px">s</span><span style=";font-family:Calibri;font-size:12px">, respectively, so that the final concentration of arsenic was (0, 5ppb, 10ppb, 50ppb, 100ppb, 500ppb, 1ppm, 5ppm, 10ppm, 50ppm and 100ppm). Samples were taken at different time points of 0h, 1h, 2h…8h and overnight (16h) at one hour interval and </span><span style=";font-family:Calibri;font-size:12px">OD600 value were measured with a Multiskan Spectrum Microplate Reader. </span></p><p><span style=";font-family:Calibri;font-size:14px">&nbsp;</span></p><p><span style=";font-family:Calibri;font-size:14px">&nbsp;</span></p><p><strong><span style="font-family: 'Calibri Light';font-size: 19px;background: rgb(255, 255, 255)">2</span></strong><strong><span style="font-family: 宋体;font-size: 19px;background: rgb(255, 255, 255)">. </span></strong><strong><span style="font-family: 'Calibri Light';font-size: 19px;background: rgb(255, 255, 255)">GFP expression of E.coli under the control of J33201 induced by different concentration of arsenite (As</span></strong><strong><sup><span style="font-family: 'Calibri Light';font-size: 19px;vertical-align: super;background: rgb(255, 255, 255)">3+</span></sup></strong><strong><span style="font-family: 'Calibri Light';font-size: 19px;background: rgb(255, 255, 255)">)</span></strong></p><p><span style=";font-family:宋体;font-size:16px">As</span><span style=";font-family:Calibri;font-size:16px">&nbsp;shown in Fig.2, our constructed biosensor based on J33201 part is very sensitive to arsenite(As</span><sup><span style=";font-family:Calibri;font-size:16px;vertical-align:super">3+</span></sup><span style=";font-family:Calibri;font-size:16px">) and the fluoresence signal can be detected at 50ppb arsenite. Significant dose-dependent effect was observed at the range of 50ppb-1ppm arsenite. The threshold of this biosensor is 10ppm. As the arsenite concentration increased to 50ppm, it does’t work due to the toxicity to cell at this level.</span></p><p><span style=";font-family:宋体;font-size:14px">Fig.2a</span></p><p style="text-align:center"><img alt="Characterization(图2)" width="554" height="372" src="https://2019.igem.org/wiki/images/b/b5/T--Worldshaper-wuhan--GFP1-fig2.jpeg" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"></p><p><span style=";font-family:Calibri;font-size:14px">&nbsp;</span><br></p><p><span style=";font-family:宋体;font-size:14px">Fig</span><span style=";font-family:Calibri;font-size:14px">.2b</span></p><p style="text-align:center"><img alt="Characterization(图3)" width="373" height="110" src="https://2019.igem.org/wiki/images/3/33/T--Worldshaper-wuhan--GFP1-fig3.jpeg" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"></p><p><span style=";font-family:Calibri;font-size:14px">&nbsp;</span><br></p><p><strong><span style="font-family: Calibri;font-size: 12px">Fig.2 GFP expression of E.coli under the control of J33201 in different concentration of arsenite (As</span></strong><strong><sup><span style="font-family: Calibri;font-size: 12px;vertical-align: super">3+</span></sup></strong><strong><span style="font-family: Calibri;font-size: 12px">)</span></strong><span style=";font-family:Calibri;font-size:12px">. </span><strong><span style="font-family: Calibri;font-size: 12px">(a)</span></strong><span style=";font-family:Calibri;font-size:12px">&nbsp;Overnight cultured bacterial solution was inoculated in LB broth containing chloramphenicol at 1:50 to expand the culture, and the experiment was started when OD600 reached 0.4-0.6. Different concentrations of arsenite solutions were added into the test tube, respectively, so that the final concentration of arsenic was (0,5ppb, 10ppb, 50ppb, 100ppb, 500ppb, 1ppm, 5ppm, 10ppm, 50ppm, 100ppm). Samples were taken at overnight (16h). GFP fluorescence intensity (485 nm excitation/ 528 nm emission) and OD600 value were measured at the same time. &nbsp;</span><strong><span style="font-family: Calibri;font-size: 12px">(b)</span></strong><span style=";font-family:Calibri;font-size:12px">The bacteria were centrifuged, and the pellets were was observed and photographed under Blue Light Gel Imager.</span></p><p><span style=";font-family:Calibri;font-size:14px">&nbsp;</span></p><p><span style=";font-family:Calibri;font-size:16px">As it is shown in </span><span style=";font-family:宋体;font-size:16px">F</span><span style=";font-family:Calibri;font-size:16px">ig.3</span><span style=";font-family:宋体;font-size:16px">, </span><span style=";font-family:Calibri;font-size:16px">our constructed biosensor by J33201 reacted rapidly in arsenite solution. When the arsenite is added for 4 hours, the corresponding fluorescence signal can be detected at 100ppb. As time increased, weak signal can be detected at 50ppb, indicating that the biosensor can work within 4 hours. </span></p><p style="text-align:center"><img alt="Characterization(图4)" width="554" height="372" src="https://2019.igem.org/wiki/images/3/36/T--Worldshaper-wuhan--GFP2-fig4.jpeg" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"></p><p><span style=";font-family:Calibri;font-size:14px">&nbsp;</span><br></p><p><strong><span style="font-family: Calibri;font-size: 12px">Fig.3 GFP expression of E.coli in different concentration of arsenite (As3+) at different time points under the control of J33201.</span></strong><span style=";font-family:Calibri;font-size:12px">&nbsp;Overnight cultured bacterial solution was inoculated in LB broth containing chloramphenicol resistance at 1:50 to expand the culture, and the experiment was started when OD600 reached 0.4-0.6.</span><span style=";font-family:Calibri;font-size:12px">&nbsp;</span><span style=";font-family:Calibri;font-size:12px">Different concentrations of arsenite solutions were added into the test tube, respectively, so that the final concentration of arsenic was (0, 5ppb, 10ppb, 50ppb, 100ppb, 500ppb, 1ppm, 5ppm, 10ppm, 50ppm, 100ppm). </span><span style=";font-family:Calibri;font-size:12px">Samples were taken at different time points of 0h, 2h, 4h, 6h and overnight (16h).</span><span style=";font-family:Calibri;font-size:12px">&nbsp;GFP fluorescence intensity (485 nm excitation/ 528 nm emission) and OD600 value were measured at the same time</span></p><p><span style=";font-family:Calibri">&nbsp;</span></p><p><span style=";font-family:Calibri;font-size:16px">To summary, our biosensor constructed on the basis of J33201 is sens</span><span style=";font-family:宋体;font-size:16px">itive and fast. </span><span style=";font-family:Calibri;font-size:16px">The biosensor can detect 100ppb-10ppm arsenite (As3+) in water within 4 hours, and the minimum detection limit can reach 50ppb with the extension of time.</span></p><p><br></p>
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Latest revision as of 00:43, 22 October 2019

Description_worldshaper-wuhan

Measurement

Measurement

Aim of experiment 

Based on J33201 part, a biosensor pSB1C3-pArsR-RBS-GFP(K3153000) was constructed to detect arsenite(As3+) in water and contributed to this well-characterized part. 

Methods

1. Growth curve of E.coli containing part K577881 in different concentrations of arsenite(As3+)

Constructed plasmid pSB1C3-pArsR-GFP containing J33201 part was transformed into E.coli DH5α strain. Single colony was selected to inoculate LB broth containing chloramphenicol and cultured overnight. Then overnight culture was inoculated in the fresh LB medium containing 34 μg/ml chloramphenicol at a ratio of 1:100, mixed well and divided into tubes. Different concentrations of arsenite (As3+) solutions were added into the test tubes, respectively. The final concentration of arsenic was (0, 5ppb, 10ppb, 50ppb, 100ppb, 500ppb, 1ppm, 5ppm, 10ppm, 50ppm and 100ppm). Samples were collected at different time points of 0 h, 1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 7 h, 8 h and 16 h. OD600 values were measured by the Multiskan Spectrum Microplate Reader.

2. GFP expression of E.coli under the control of J33201 induced by different concentration of arsenite (As3+)

Overnight cultured bacterial solution was inoculated in LB broth containing chloramphenicol at 1:50 to expand the culture. When OD600 reached 0.4-0.6, experiments were performed . Different concentrations of arsenite were added into the test tube, respectively. The final concentration of arsenic was 0, 5 ppb, 10 ppb, 50 ppb, 100 ppb, 500 ppb, 1 ppm, 5 ppm, 10 ppm, 50 ppm, 100 ppm.

(1) Samples were collected at 16 h. GFP fluorescence intensity (485 nm excitation/ 528 nm emission) and OD600 value were measured at the same time. The bacteria were centrifuged, and the pellets were was observed and photographed under Blue Light Gel Imager.

(2)Samples were collected at different time points of 0 h, 2 h, 4 h, 6 h and 16 h. GFP fluorescence intensity (485 nm excitation/ 528 nm emission) and OD600 value were measured at the same time.

 

Results

Fig.1 showed that bacteria growth was not affected from 5 ppb to 10 ppm of arsenite solution (As3+). However, when the concentration of arsenite increased to 50 ppm, the growth of the bacteria was seriously inhibited, suggesting the toxictity to cells at this concentration.

Characterization(图1)

 

Fig.1 Growth curve of E.coli in different concentration of arsenic (As3+).

 

As shown in Fig.2, our biosensor based on J33201 part is very sensitive to arsenite(As3+) and the fluoresence signal was be detected at 50 ppb arsenite. Significant dose-dependent effect was observed from 50 ppb to 1 ppm arsenite. The threshold of this biosensor is 10 ppm. As the arsenite concentration increased to 50 ppm, it does’t work because of the toxicity to cell at thisconcentration.

Characterization(图2)

 

(Fig.2a)

Characterization(图3)

 

(Fig.2b)

Fig.2 GFP expression of E.coli under the control of J33201 in different concentration of arsenite (As3+).

 

As shown in Fig.3, our biosensor by J33201 reacted rapidly in arsenite solution. After 4 h following arsenite treated, the corresponding fluorescence signal was detected at 100 ppb. As time increases, a weak signal can be detected at 50ppb, indicating that the biosensor can work within 4 hours.

Characterization(图4)

 

Fig.3 GFP expression of E.coli in different concentration of arsenite (As3+) at different time points under the control of J33201.

 

Summary

our biosensor constructed on the basis of J33201 is sensitive and fast. The biosensor can detect 100ppb-10ppm arsenite (As3+) in water within 4 hours. With the extension of time, the minimum detection limit can reach 50 ppb.

Reference

Anal Bioanal Chem. 2011 May;400(4):1031-9. Epub 2011 Mar 27.