Difference between revisions of "Team:NJU-China/Notebook/Calendar"

 
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    color: #726f77;
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    max-width: 1100px;
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/*h1, h2, h3, h4 {
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    font-family: "Dosis", arial, tahoma, verdana;
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    font-weight: 600;
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#timeline {
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<head>
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     width: 90%;
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     <meta charset="UTF-8">
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    margin: 30px auto;
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     <meta http-equiv="Content-Type" content="text/html;charset=utf-8">
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    position: relative;
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     <meta name="viewport" content="width=device-width, initial-scale=1.0, minimum-scale=1.0">
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     -webkit-transition: all 0.4s ease;
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     <!--Import materialize.css-->
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     -moz-transition: all 0.4s ease;
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     <link rel="stylesheet" href="https://2016.igem.org/Team:NJU-China/mdlib/ghpages?action=raw&amp;ctype=text/css" media="screen,projection">
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     -ms-transition: all 0.4s ease;
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     <!--Import Google Icon Font-->
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     transition: all 0.4s ease;
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     <link href="https://2016.igem.org/Team:NJU-China/mdlib/mdiconcss?action=raw&amp;ctype=text/css" rel="stylesheet">
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     <link href="https://2016.igem.org/Team:NJU-China/mdlib/customedcss?action=raw&amp;ctype=text/css" rel="stylesheet">
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#timeline:before {
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     <link href="https://2016.igem.org/Team:NJU-China/mdlib/calendarcss?action=raw&amp;ctype=text/css" rel="stylesheet">
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    content: "";
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</head>
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     width: 3px;
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−
     height: 100%;
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<body>
−
     background: #ee4d4d;
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     <header>
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     left: 50%;
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        <nav class="top-nav">
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     top: 0;
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            <div class="container">
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    position: absolute;
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                <div class="nav-wrapper"><a class="page-title">Calendar</a></div>
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}
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            </div>
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#timeline:after {
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        </nav>
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    content: "";
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        <div class="container">
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    clear: both;
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            <a href="#" data-activates="nav-mobile" class="button-collapse top-nav full hide-on-large-only"> <i class="material-icons">menu</i> </a>
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    display: table;
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        </div>
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    width: 100%}
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        <ul id="nav-mobile" class="side-nav fixed">
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#timeline .timeline-item {
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            <li class="logo">
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    margin-bottom: 50px;
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                <a href="https://2016.igem.org/Team:NJU-China"><img class="background responsive-img" src="https://static.igem.org/mediawiki/2016/c/c8/NJU_China_2016_iGEM_logo.png" alt="NJU-China LOGO"></a>
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    position: relative;
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            </li>
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}
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            <li class="bold"><a href="https://2016.igem.org/Team:NJU-China/Background" class="waves-effect waves-teal">Background</a></li>
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#timeline .timeline-item .timeline-icon {
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            <li class="bold no-padding">
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    background: #ee4d4d;
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                <ul class="collapsible collapsible-accordion">
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    width: 50px;
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                    <li class="bold"> <a class="collapsible-header waves-effect waves-teal">Project Design</a>
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    height: 50px;
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                        <div class="collapsible-body">
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    position: absolute;
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                            <ul>
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    top: 0;
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Project_Design">RNAi Module</a></li>
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    left: 50%;
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Project_Design#Targeting_Module">Targeting Module</a></li>
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    overflow: hidden;
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Project_Design#Assembly">Assembly</a></li>
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    margin-left: -23px;
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                            </ul>
−
    -webkit-border-radius: 50%;
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                        </div>
−
    -moz-border-radius: 50%;
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                    </li>
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    -ms-border-radius: 50%;
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                </ul>
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    border-radius: 50%}
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            </li>
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#timeline .timeline-item .timeline-icon img {
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            <li class="bold"><a href="https://2016.igem.org/Team:NJU-China/Modeling" class="waves-effect waves-teal">Modeling</a></li>
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    position: relative;
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            <li class="bold no-padding">
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    top: 14px;
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                <ul class="collapsible collapsible-accordion">
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    left: 14px;
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                    <li class="bold"> <a class="collapsible-header waves-effect waves-teal">Results</a>
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}
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                        <div class="collapsible-body">
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#timeline .timeline-item .timeline-content {
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                            <ul>
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     width: 45%;
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Results">Parts</a></li>
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    background: #fff;
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Results#Validations">Validations</a></li>
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    padding: 20px;
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Results#Safety">Safety</a></li>
−
    -webkit-box-shadow: 0 3px 0 rgba(0, 0, 0, 0.1);
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Results#Conclusions">Conclusions</a></li>
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    -moz-box-shadow: 0 3px 0 rgba(0, 0, 0, 0.1);
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                            </ul>
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                    </li>
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    -webkit-border-radius: 5px;
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                </ul>
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    -moz-border-radius: 5px;
+
            </li>
−
    -ms-border-radius: 5px;
+
            <li class="bold"><a href="https://2016.igem.org/Team:NJU-China/Human_Practices" class="waves-effect waves-teal">Human Practices</a></li>
−
    border-radius: 5px;
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            <li class="bold"><a href="https://2016.igem.org/Team:NJU-China/Team" class="waves-effect waves-teal">Team</a></li>
−
    -webkit-transition: all 0.3s ease;
+
            <li class="bold"><a href="https://2016.igem.org/Team:NJU-China/Attributions" class="waves-effect waves-teal">Attributions</a></li>
−
    -moz-transition: all 0.3s ease;
+
            <li class="bold"><a href="https://2016.igem.org/Team:NJU-China/Collaborations" class="waves-effect waves-teal">Collaborations</a></li>
−
    -ms-transition: all 0.3s ease;
+
            <li class="bold no-padding">
−
    transition: all 0.3s ease;
+
                <ul class="collapsible collapsible-accordion">
−
}
+
                    <li class="bold"> <a class="collapsible-header waves-effect waves-teal">Notebook</a>
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#timeline .timeline-item .timeline-content h2 {
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                        <div class="collapsible-body">
−
    padding: 15px;
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                            <ul>
−
    background: #ee4d4d;
+
                                <li><a href="https://2016.igem.org/Team:NJU-China/Notebook/Calendar">Calendar</a></li>
−
    color: #fff;
+
                                <li><a href="https://2016.igem.org/Team:NJU-China/Notebook/Protocol">Protocol</a></li>
−
    margin: -20px -20px 0 -20px;
+
                            </ul>
−
    font-weight: 500;
+
                        </div>
−
    -webkit-border-radius: 3px 3px 0 0;
+
                    </li>
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    -moz-border-radius: 3px 3px 0 0;
+
                </ul>
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    -ms-border-radius: 3px 3px 0 0;
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            </li>
−
    border-radius: 3px 3px 0 0;
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        </ul>
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}
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    </header>
−
#timeline .timeline-item .timeline-content:before {
+
    <!-- Begin page content -->
−
    content: '';
+
     <main>
−
    position: absolute;
+
        <div class="fixed-action-btn" style="bottom: 45px; right: 24px;">
−
    left: 45%;
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            <a class="btn-floating btn-large red">
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                <i class="large material-icons">add</i>
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    height: 0;
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            <ul>
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    border-top: 7px solid transparent;
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                <li onclick="$('html, body').animate({ scrollTop: 0 }, 'fast');"><a class="btn-floating green tooltipped" data-position="left" data-delay="50" data-tooltip="Scroll to Top"><i class="material-icons">arrow_upward</i></a></li>
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    border-bottom: 7px solid transparent;
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                <li onclick="$('html, body').animate({scrollTop: $('html').height()-$(window).height()}, 'fast')"><a class="btn-floating yellow darken-2 tooltipped" data-position="left" data-delay="50" data-tooltip="Scroll to Bottom"><i class="material-icons">arrow_downward</i></a></li>
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}
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                <li><a class="btn-floating blue button-collapse hide-on-large-only tooltipped" data-position="left" data-delay="50" data-tooltip="Show Navigation" data-activates="nav-mobile"><i class="material-icons">menu</i></a></li>
−
#timeline .timeline-item .timeline-content.right {
+
            </ul>
−
    float: right;
+
        </div>
−
}
+
        <div class="container">
−
#timeline .timeline-item .timeline-content.right:before {
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            <div id="timeline">
−
    content: '';
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                <div class="timeline-item">
−
    right: 45%;
+
                    <div class="timeline-icon">
−
    left: inherit;
+
                        <i class="material-icons">timeline</i>
−
    border-left: 0;
+
                    </div>
−
    border-right: 7px solid #ee4d4d;
+
                    <div class="timeline-content">
−
}
+
                        <h4>Week 1 (05/22/2016-05/28/2016)</h4>
−
/*.btn {
+
                        <p>Learn laboratory safety regulations and lab skills like PCR, restriction enzyme digestion, electrophoresis, fragment ligation, etc.</p>
−
    padding: 5px 15px;
+
                        <p>Acquire iRGD-Lamp2b fusion protein sequence from GenScript, then try to construct plasmid (pcDNA6.2) ---Failed</p>
−
    text-decoration: none;
+
                    </div>
−
    background: transparent;
+
                </div>
−
    border: 2px solid #f27c7c;
+
                <div class="timeline-item">
−
    color: #f27c7c;
+
                    <div class="timeline-icon">
−
    display: inline-block;
+
                        <i class="material-icons">timeline</i>
−
    position: relative;
+
                    </div>
−
    text-transform: uppercase;
+
                    <div class="timeline-content right">
−
    font-size: 12px;
+
                        <h4>Week 2 (05/29/2016-06/04/2016)</h4>
−
    -webkit-border-radius: 5px;
+
                        <p>PCR, Plasmid construction using iRGD-Lamp2b sequence and pSB1C3 backbone.---Failed</p>
−
    -moz-border-radius: 5px;
+
                        <p>Plasmid construction using iRGD-Lamp2b sequence and pSB1C3 backbone.---Failed</p>
−
    -ms-border-radius: 5px;
+
                        <p>Learn cellular experiment skills. (cell culture & transfection)</p>
−
    border-radius: 5px;
+
                        <p>HEK293, A549-Luc, A549 cell lines recovery.</p>
−
    -webkit-transition: background 0.3s ease;
+
                    </div>
−
    -moz-transition: background 0.3s ease;
+
                </div>
−
    -ms-transition: background 0.3s ease;
+
                <div class="timeline-item">
−
    transition: background 0.3s ease;
+
                    <div class="timeline-icon">
−
    -webkit-box-shadow: 2px 2px 0 #f27c7c;
+
                        <i class="material-icons">timeline</i>
−
    -moz-box-shadow: 2px 2px 0 #f27c7c;
+
                    </div>
−
    -ms-box-shadow: 2px 2px 0 #f27c7c;
+
                    <div class="timeline-content">
−
    box-shadow: 2px 2px 0 #f27c7c;
+
                        <h4>Week 3 (06/05/2016-06/11/2016)</h4>
−
}
+
                        <p>Plasmid construction using iRGD-Lamp2b sequence and pSB1C3 backbone.---Succeed</p>
−
.btn:hover {
+
                        <p>Learn western blot protocols.</p>
−
    box-shadow: none;
+
                    </div>
−
    top: 2px;
+
                </div>
−
    left: 2px;
+
                <div class="timeline-item">
−
    -webkit-box-shadow: 2px 2px 0 transparent;
+
                    <div class="timeline-icon">
−
    -moz-box-shadow: 2px 2px 0 transparent;
+
                        <i class="material-icons">timeline</i>
−
    -ms-box-shadow: 2px 2px 0 transparent;
+
                    </div>
−
    box-shadow: 2px 2px 0 transparent;
+
                    <div class="timeline-content right">
−
}*/
+
                        <h4>Week 4 (06/12/2016-06/18/2016)</h4>
−
@media screen and (max-width: 768px) {
+
                        <p>Test software written by team SYSU-Software . Design siRNAs.</p>
−
    #timeline {
+
                        <p>Order siRNA from GenScript.</p>
−
    margin: 30px;
+
                    </div>
−
    padding: 0;
+
                </div>
−
}
+
                <div class="timeline-item">
−
#timeline:before {
+
                    <div class="timeline-icon">
−
    left: 0;
+
                        <i class="material-icons">timeline</i>
−
}
+
                    </div>
−
#timeline .timeline-item .timeline-content {
+
                    <div class="timeline-content">
−
    width: 90%;
+
                        <h4>Week 5 (06/19/2016-06/25/2016)</h4>
−
    float: right;
+
                        <p>Receive siRNA from GenScript, Test RNA interference efficiency by cell transfection and western blotting.--- Failed</p>
−
}
+
                        <p>Test KRAS expression level in which cell line using transfection and western blotting.---Succeed</p>
−
#timeline .timeline-item .timeline-content:before, #timeline .timeline-item .timeline-content.right:before {
+
                        <p>Abandon cells contaminated with bacteria. Disinfection and sterilization in cell incubators.</p>
−
     left: 10%;
+
                        <p>Learn protocols for FCFM, ultra-high-speed centrifuge, in-vivo imaging.</p>
−
     margin-left: -6px;
+
                        <p>Obtain access for State Key Laboratory of Pharmaceutical Biotechnology.</p>
−
     border-left: 0;
+
                    </div>
−
     border-right: 7px solid #ee4d4d;
+
                </div>
−
}
+
                <div class="timeline-item">
−
#timeline .timeline-item .timeline-icon {
+
                    <div class="timeline-icon">
−
     left: 0;
+
                        <i class="material-icons">timeline</i>
−
}
+
                    </div>
−
}
+
                    <div class="timeline-content right">
 +
                        <h4>Week 6 (06/26/2016-07/02/2016)</h4>
 +
                        <p>A549-Luc cell line recovery and culture. Preparation for animal modeling.</p>
 +
                        <p>Design shRNA based on validated siRNA sequence. Send our designed sequence to GenScript for synthesis.</p>
 +
                        <p>Successfully loading our exosomes with iRGD and siRNA for the first time. Send exosome samples to National University of Defense Technology for electron microscopy assay and Nanosight.</p>
 +
                    </div>
 +
                </div>
 +
                <div class="timeline-item">
 +
                    <div class="timeline-icon">
 +
                        <i class="material-icons">timeline</i>
 +
                    </div>
 +
                    <div class="timeline-content">
 +
                        <h4>Week 7 (07/03/2016-07/09/2016)</h4>
 +
                        <p>One-week suspension for thoroughly disinfection and sterilization.</p>
 +
                    </div>
 +
                </div>
 +
                <div class="timeline-item">
 +
                    <div class="timeline-icon">
 +
                        <i class="material-icons">timeline</i>
 +
                    </div>
 +
                    <div class="timeline-content right">
 +
                        <h4>Week 8 (07/10/2016-07/16/2016)</h4>
 +
                        <p>Receive feedbacks from National University of Defense Technology.</p>
 +
                        <p>HEK293, A549-Luc, A549 cell lines recovery.</p>
 +
                        <p>Detecting KRAS expression level using western blotting and qPCR.</p>
 +
                        <p>A549-Luc cell proliferation for mouse tumor model construction.</p>
 +
                        <p>Verify the effect of exosome on cell proliferation of A549</p>
 +
                    </div>
 +
                </div>
 +
                <div class="timeline-item">
 +
                    <div class="timeline-icon">
 +
                        <i class="material-icons">timeline</i>
 +
                    </div>
 +
                    <div class="timeline-content">
 +
                        <h4>Week 9 (07/17/2016-07/23/2016)</h4>
 +
                        <p>A549-Luc cell proliferation in preparation for tumor implantation</p>
 +
                        <p>HEK293 cell proliferation for mass-production of exosomes</p>
 +
                        <p>Order nude mice from Model Animal Research Center of Nanjing University and inoculate A549-Luc subcutaneously into nude mice to develop implant-tumor.</p>
 +
                        <p>Receive two single strands of RNA containing KRAS shRNA sequence from Genscript</p>
 +
                        <p>Learn work standards of animal experiments. Obtain permission for animal experiment in School of Life Science, NJU</p>
 +
                    </div>
 +
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 +
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 +
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 +
                        <h4>Week 10 (07/24/2016-07/30/2016)</h4>
 +
                        <p>Abandon HEK293 cells with obviously slower cell proliferation and changing characteristics. Recover HEK293 cells.</p>
 +
                        <p>Anneal two single strands of shRNA into segments. Try to insert these fragments into pSB1C3 plasmid vectors. ---Failed</p>
 +
                    </div>
 +
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 +
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 +
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 +
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 +
                        <h4>Week 11 (07/31/2016-08/06/2016)</h4>
 +
                        <p>Detect endotoxin level of exosome</p>
 +
                        <p>HEK293 cell proliferation</p>
 +
                        <p>Retry to anneal two single strands of shRNA into segments and insert them into pSB1C3 plasmid vectors. ---succeed</p>
 +
                    </div>
 +
                </div>
 +
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 +
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 +
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 +
                        <h4>Week 12 (08/07/2016-08/13/2016)</h4>
 +
                        <p>Perform in vivo imaging on nude mice for experimental model and most tumor implantations were successful</p>
 +
                        <p>Randomly divide the mouse model into two groups</p>
 +
                        <p>HEK293 cell proliferation</p>
 +
                        <p>HEK293 cells amplification rates decrease and cells are thawed</p>
 +
                    </div>
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 +
                        <h4>Week 13 (08/14/2016-08/20/2016)</h4>
 +
                        <p>HEK293 cell proliferation</p>
 +
                        <p>HEK293 cell are abandoned for decreased viability.</p>
 +
                    </div>
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 +
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 +
                        <h4>Week 14 (08/21/2016-08/27/2016)</h4>
 +
                        <p>HEK293 cell line recovery and proliferation.</p>
 +
                    </div>
 +
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 +
                        <h4>Week 15 (08/28/2016-09/03/2016)</h4>
 +
                        <p>HEK293 cell proliferation</p>
 +
                    </div>
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 +
                        <h4>Week16 (09/04/2016-09/10/2016)</h4>
 +
                        <p>Obtain enough HEK293 cells. Transfect HEK293 cells with KRAS siRNA. Isolate exosomes from cell culture.</p>
 +
                        <p>Mouse tail vein injection</p>
 +
                    </div>
 +
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 +
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 +
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 +
                        <h4>Week17 (09/11/2016-09/17/2016)</h4>
 +
                        <p>HEK 293 cell proliferation. Transfect these cells and collect exosome</p>
 +
                        <p>Mouse tail vein injection</p>
 +
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                        <h4>Week18 (09/18/2016-09/24/2016)</h4>
 +
                        <p>Stop mouse tail vein injection ahead of schedule since time was limited</p>
 +
                        <p>Detect the effects of our exosome through in vivo imaging</p>
 +
                        <p>Sacrifice these nude mice, collect their tumor tissues and record data</p>
 +
                    </div>
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 +
                        <h4>Week19 (09/25/2016-10/01/2016)</h4>
 +
                        <p>Detect the expression level of KRAS mRNA and K-ras protein in these tumor tissues</p>
 +
                        <p>Frozen sectioning using tumor tissues. Observe relevant pathological characteristics.</p>
 +
                    </div>
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Latest revision as of 02:53, 20 October 2016

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Week 1 (05/22/2016-05/28/2016)

Learn laboratory safety regulations and lab skills like PCR, restriction enzyme digestion, electrophoresis, fragment ligation, etc.

Acquire iRGD-Lamp2b fusion protein sequence from GenScript, then try to construct plasmid (pcDNA6.2) ---Failed

timeline

Week 2 (05/29/2016-06/04/2016)

PCR, Plasmid construction using iRGD-Lamp2b sequence and pSB1C3 backbone.---Failed

Plasmid construction using iRGD-Lamp2b sequence and pSB1C3 backbone.---Failed

Learn cellular experiment skills. (cell culture & transfection)

HEK293, A549-Luc, A549 cell lines recovery.

timeline

Week 3 (06/05/2016-06/11/2016)

Plasmid construction using iRGD-Lamp2b sequence and pSB1C3 backbone.---Succeed

Learn western blot protocols.

timeline

Week 4 (06/12/2016-06/18/2016)

Test software written by team SYSU-Software . Design siRNAs.

Order siRNA from GenScript.

timeline

Week 5 (06/19/2016-06/25/2016)

Receive siRNA from GenScript, Test RNA interference efficiency by cell transfection and western blotting.--- Failed

Test KRAS expression level in which cell line using transfection and western blotting.---Succeed

Abandon cells contaminated with bacteria. Disinfection and sterilization in cell incubators.

Learn protocols for FCFM, ultra-high-speed centrifuge, in-vivo imaging.

Obtain access for State Key Laboratory of Pharmaceutical Biotechnology.

timeline

Week 6 (06/26/2016-07/02/2016)

A549-Luc cell line recovery and culture. Preparation for animal modeling.

Design shRNA based on validated siRNA sequence. Send our designed sequence to GenScript for synthesis.

Successfully loading our exosomes with iRGD and siRNA for the first time. Send exosome samples to National University of Defense Technology for electron microscopy assay and Nanosight.

timeline

Week 7 (07/03/2016-07/09/2016)

One-week suspension for thoroughly disinfection and sterilization.

timeline

Week 8 (07/10/2016-07/16/2016)

Receive feedbacks from National University of Defense Technology.

HEK293, A549-Luc, A549 cell lines recovery.

Detecting KRAS expression level using western blotting and qPCR.

A549-Luc cell proliferation for mouse tumor model construction.

Verify the effect of exosome on cell proliferation of A549

timeline

Week 9 (07/17/2016-07/23/2016)

A549-Luc cell proliferation in preparation for tumor implantation

HEK293 cell proliferation for mass-production of exosomes

Order nude mice from Model Animal Research Center of Nanjing University and inoculate A549-Luc subcutaneously into nude mice to develop implant-tumor.

Receive two single strands of RNA containing KRAS shRNA sequence from Genscript

Learn work standards of animal experiments. Obtain permission for animal experiment in School of Life Science, NJU

timeline

Week 10 (07/24/2016-07/30/2016)

Abandon HEK293 cells with obviously slower cell proliferation and changing characteristics. Recover HEK293 cells.

Anneal two single strands of shRNA into segments. Try to insert these fragments into pSB1C3 plasmid vectors. ---Failed

timeline

Week 11 (07/31/2016-08/06/2016)

Detect endotoxin level of exosome

HEK293 cell proliferation

Retry to anneal two single strands of shRNA into segments and insert them into pSB1C3 plasmid vectors. ---succeed

timeline

Week 12 (08/07/2016-08/13/2016)

Perform in vivo imaging on nude mice for experimental model and most tumor implantations were successful

Randomly divide the mouse model into two groups

HEK293 cell proliferation

HEK293 cells amplification rates decrease and cells are thawed

timeline

Week 13 (08/14/2016-08/20/2016)

HEK293 cell proliferation

HEK293 cell are abandoned for decreased viability.

timeline

Week 14 (08/21/2016-08/27/2016)

HEK293 cell line recovery and proliferation.

timeline

Week 15 (08/28/2016-09/03/2016)

HEK293 cell proliferation

timeline

Week16 (09/04/2016-09/10/2016)

Obtain enough HEK293 cells. Transfect HEK293 cells with KRAS siRNA. Isolate exosomes from cell culture.

Mouse tail vein injection

timeline

Week17 (09/11/2016-09/17/2016)

HEK 293 cell proliferation. Transfect these cells and collect exosome

Mouse tail vein injection

timeline

Week18 (09/18/2016-09/24/2016)

Stop mouse tail vein injection ahead of schedule since time was limited

Detect the effects of our exosome through in vivo imaging

Sacrifice these nude mice, collect their tumor tissues and record data

timeline

Week19 (09/25/2016-10/01/2016)

Detect the expression level of KRAS mRNA and K-ras protein in these tumor tissues

Frozen sectioning using tumor tissues. Observe relevant pathological characteristics.