Difference between revisions of "Team:NYMU-Taipei/Notebook"

 
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{{NYMU-Taipei-css}}
 
{{NYMU-Taipei-css}}
  
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<html>
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<html lang="en">
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<head>
 
<head>
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<meta charset="utf-8">
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<meta http-equiv="X-UA-Compatible" content="IE=edge">
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<meta name="viewport" content="width=device-width, initial-scale=1">
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<!-- Bootstrap -->
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<link rel="stylesheet" href="https://maxcdn.bootstrapcdn.com/bootstrap/3.3.7/css/bootstrap.min.css">
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<script src="https://ajax.googleapis.com/ajax/libs/jquery/3.2.0/jquery.min.js"></script>
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<script src="https://maxcdn.bootstrapcdn.com/bootstrap/3.3.7/js/bootstrap.min.js"></script>
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<style>
 
<style>
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body{
 
body{
 
background-color:#f6ffb2;
 
background-color:#f6ffb2;
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}
 
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/* Wrapper for notebook */
 
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.notebook_wrapper{
 
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width: 85%;
 
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display:block;
 
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float:right;
 
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font-family:'Fresca',sans-serif;
 
 
}
 
}
 
 
 
/* Content of Notebook */
 
/* Content of Notebook */
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#PCR{
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#Protocol{
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width: 85%;
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width: 81%;
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padding:2%;
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padding: 2%;
 
display:block;
 
display:block;
 
float:right;  
 
float:right;  
 
background-color:#f6ffb2;  
 
background-color:#f6ffb2;  
 
font-family:'Fresca',sans-serif;
 
font-family:'Fresca',sans-serif;
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font-size:20px;
 
}
 
}
 
 
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#OD_measurment{
 
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width: 85%;
 
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padding:2%;
 
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display:block;
 
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float:right;
 
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background-color:#f6ffb2;
 
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font-family:'Fresca',sans-serif;
 
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}
 
 
#Timeline{
 
#Timeline{
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width: 85%;
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width: 81%;
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padding:2%;
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padding: 2%;
 
display:block;
 
display:block;
 
float:right;  
 
float:right;  
 
background-color:#cee1ea;  
 
background-color:#cee1ea;  
 
font-family:'Fresca',sans-serif;
 
font-family:'Fresca',sans-serif;
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}
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font-size:18px;
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}
 
 
 
#Labnote{
 
#Labnote{
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width: 81%;
 
padding:2%;
 
padding:2%;
 
display:block;
 
display:block;
 
float:right;  
 
float:right;  
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background-color:#ffeaea;  
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background-color: #ffeaea;  
 
font-family:'Fresca',sans-serif;
 
font-family:'Fresca',sans-serif;
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font-size:20px;
 
padding-bottom:250px;
 
padding-bottom:250px;
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}
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}
 
 
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#PCR h1 {
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#Protocol h1 {
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color: #424017;
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color: #205e1a;
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font-size: 30px;
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font-size: 40px;
 
font-family: 'Acme', sans-serif;
 
font-family: 'Acme', sans-serif;
 
}
 
}
 
 
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#PCR h3 {
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#Protocol h3 {
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color: #205e1a;
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color: #3dba32;
 
font-size: 28px;
 
font-size: 28px;
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font-family: 'Francois One', sans-serif;
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font-family: 'Acme', sans-serif;
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padding-left:20px;
 
}
 
}
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#OD_measurment h3 {
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#Protocol a {
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color: #205e1a;
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color:#205e1a;
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font-size: 28px;
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font-family: 'Acme', sans-serif;
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font-family: 'Francois One', sans-serif;
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}
 
}
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#Timeline h1{
 
#Timeline h1{
 
color: #1c3e4f;
 
color: #1c3e4f;
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font-size: 30px;
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font-size: 40px;
 
font-family: 'Acme', sans-serif;
 
font-family: 'Acme', sans-serif;
 
}
 
}
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#Labnote h1{
 
#Labnote h1{
 
color: #722525;
 
color: #722525;
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font-size: 30px;
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font-size: 40px;
 
font-family: 'Acme', sans-serif;
 
font-family: 'Acme', sans-serif;
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}
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}
 
 
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/* Wrapper for the footer */
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/* Show & hide */
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.igem_2017_footer_wrapper {
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.expandable {
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width: 85%;
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/* overflow set to hidden to hide the expanded text */
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padding: 0.8% 2%;
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overflow: hidden;
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display:block;
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/* all style changes will ease-in-out for 1s */
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float: right;
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-moz-transition: all 1s ease-in-out;
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background:#f6ffb2 url('https://static.igem.org/mediawiki/2017/0/0f/T--NYMU-Taipei--footer.png') bottom no-repeat;
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-ms-transition: all 1s ease-in-out;
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height:150px;
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-o-transition: all 1s ease-in-out;
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position: absolute;
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-webkit-transition: all 1s ease-in-out;
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right: 0;
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transition: all 1s;
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bottom: 0;
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font-family:'Economica', sans-serif;  
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}
 
}
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.panel {
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background-color: #fff;
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border: 1px solid transparent;
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border-radius: 4px;
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border-color: #ddd;
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-webkit-box-shadow: 0 1px 1px rgba(0, 0, 0, .05);
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box-shadow: 0 1px 1px rgba(0, 0, 0, .05);
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padding:15px;
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padding-top:20px;
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}
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/* IF THE SCREEN IS LESS THAN 800PX */
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@media only screen and (max-width: 800px) {
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.igem_2017_footer_wrapper {width:100%; margin-left:0px;}
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#Protocol {width:96%; margin-left:50px;}
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#Timeline {width:96%; margin-left:50px;}
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#Labnote {width:96%; margin-left:50px;}
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}
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</style>
 
</style>
 
 
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</head>
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<script>
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function toggleHeight1(e, maxHeight) {
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e = document.getElementById("s1"); // e = the gray div
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if(e.style.height != '24px') {
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function toggleHeight2(e, maxHeight) {
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e = document.getElementById("s2"); // e = the gray div
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function toggleHeight3(e, maxHeight) {
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e = document.getElementById("s3"); // e = the gray div
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if(e.style.height != '24px') {
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e.style.height = maxHeight + 'px';
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function toggleHeight4(e, maxHeight) {
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e = document.getElementById("s4"); // e = the gray div
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<body data-spy="scroll" data-target=".navbar" data-offset="50">
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function toggleHeight5(e, maxHeight) {
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e = document.getElementById("s5"); // e = the gray div
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e.style.height = maxHeight + 'px';
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</script>
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</head>
 
 
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<body>
 
 
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<!--<div class = "notebook_wrapper"> -->
 
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<nav class="navbar navbar-inverse navbar-fixed-top" role="navigation" style="width:85%;float:right;display:block">
 
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<div class="container-fluid"  >
 
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<div class="navbar-header">
 
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<button type="button" class="navbar-toggle" data-toggle="collapse" data-target="#myNavbar">
 
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<span class="icon-bar"></span>
 
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<span class="icon-bar"></span>
 
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<span class="icon-bar"></span>                       
 
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</button>
 
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<a class="navbar-brand" href="#">Notebook</a>
 
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</div>
 
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<div class="collapse navbar-collapse">
 
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<ul class="nav navbar-nav">
 
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<li class="dropdown">
 
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<a href="#" id="navbarDrop1" class="dropdown-toggle"
 
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data-toggle="dropdown">Protocol
 
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<b class="caret"></b>
 
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</a>
 
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<ul class="dropdown-menu" role="menu" aria-labelledby="navbarDrop1">
 
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<li><a href="#PCR" tabindex="-1">PCR</a></li>
 
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<li><a href="#OD_measurment" tabindex="-1">OD measurment</a></li>
 
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</ul>
 
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</li>
 
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<li>
 
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<a href="#Timeline">Timeline</a>
 
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</li>
 
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<li>
 
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<a href="#Labnote">Labnote</a>
 
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</li>
 
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</ul>
 
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</div>
 
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</div>
 
 
 
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</nav>
 
−
 
 
 
 +
 +
<div id="Timeline" >
 +
                        <center>
 +
<h3></h3>
 +
<img src="https://static.igem.org/mediawiki/2017/f/f9/T--NYMU-Taipei--notebook.jpg"
 +
style="width:50%;border-radius:50%;border:5px solid #f6ffb2;">
 +
</center>
 +
<h3></h3>
 +
<h3></h3>
 +
<h1>Timeline</h1>
 +
<p></p>
 
 
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<div id="PCR" class="container-fluid">
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<div class='panel'>
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<h1>Protocol</h1>
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<div id="s1" class="expandable" style='height: 24px;padding-top:5px;'>
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<h3>PCR</h3><a herf="">Download the file</a>
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<a href="#!" onclick="toggleHeight1(this, 270); return false"
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<p>Will soon be uploaded...</p>
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style="font-family: 'Acme', sans-serif;font-size:28px;color: #479fcc;height: 20px;">
 +
June, 2017
 +
</a>
 +
<p>
 +
<br>
 +
<b>2017-06-19</b>&nbsp;&nbsp; prepare stock1 and stock5<br>
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<b>2017-06-20</b>&nbsp;&nbsp; (1) reprepare stock5<br>
 +
　　　　　(2) prepare BG11 medium<br>
 +
　　　　　(3) unfreeze <i>Synechocystis</i> sp. PCC6803 and start to cultivate in BG11<br>
 +
<b>2017-06-29</b>&nbsp;&nbsp; (1) prepare Pseudomonas aeruginosa growing medium<br>
 +
　　　　　(2) activate Pseudomonas aeruginosa and start to cultivate in Petri dish<br>
 +
　　　　　(3) subculture <i>Synechocystis</i> sp. PCC6803
 +
</p>
 +
 +
<center>
 +
<a href="#!" onclick="toggleHeight1(this, 270);" style='font-size:20px;color:#2c498c'>
 +
click to close
 +
</a>
 +
</center>
 
</div>
 
</div>
−
<div id="OD_measurment" class="container-fluid">
 
−
<h3>OD measurment</h3><a herf="">Download the file</a>
 
−
<p>Will soon be uploaded...</p>
 
 
</div>
 
</div>
−
+
−
<div id="Timeline" class="container-fluid">
+
<div class='panel'>
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<h1>Timeline</h1> <a herf="">Download the file</a>
+
<div id="s2" class="expandable" style='height: 24px;padding-top:5px;'>
−
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<a href="#!" onclick="toggleHeight2(this, 1440); return false"
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style="font-family: 'Acme', sans-serif;font-size:28px;color: #479fcc;height: 20px;">
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<div class="panel-group" id="accordion" role="tablist" aria-multiselectable="true">
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July, 2017
 +
</a>
 +
<p>
 +
<br>
 +
<b>2017-07-02</b>&nbsp;&nbsp; extract <i>Pseudomonas aeruginosa</i> plasmid and freeze into refrigerator<br>
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<b>2017-07-03</b>&nbsp;&nbsp; proliferate <i>Pseudomonas aeruginosa</i> and freeze into refrigerator<br>
 +
<b>2017-07-04</b>&nbsp;&nbsp; (1) prepare BG11 medium<br>
 +
　　　　　(2) proliferate <i>Chlorella vulgaris</i> in Petri dish and centrifuge tube<br>
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<b>2017-07-05 to 07-10</b>&nbsp;&nbsp; cultivate <i>Synechocystis</i> sp. PCC6803 and <i>Chlorella vulgaris</i> in incubator<br>
 +
<b>2017-07-11</b>&nbsp;&nbsp;&nbsp;&nbsp; (1) extract <i>Synechocystis</i> sp. PCC6803 plasmid<br>
 +
　　　　　(2) prepare LB containing ampicillin and chloramphenicol and prepare Petri dish<br>
 +
<b>2017-07-12</b>&nbsp;&nbsp; transform BBa_K112806, BBa_K737007, BBa_K112000, BBa_J04450, BBa_K1356003, BBa_K1356004, and BBa_K356005 into competent cell<br>
 +
<b>2017-07-13</b>&nbsp;&nbsp;&nbsp; (1) proliferate 07-12 transformed E.coli<br>
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　　　　　(2) proliferate <i>Chlorella vulgaris</i><br>
 +
　　　　　(3) proliferate <i>Synechocystis</i> sp. PCC6803<br>
 +
　　　　　(4) activate <i>Synechococcus elongatus</i> PCC7942<br>
 +
<b>2017-07-14</b>&nbsp;&nbsp;&nbsp; (1)extract 07-13 proliferated E.coli plasmid<br>
 +
　　　　　(2) digestion: BBa_K112806, BBa_K737007, BBa_K112000, BBa_J04450, BBa_K1356003, BBa_K1356004, and BBa_K1356005<br>
 +
　　　　　　(check the correction of transformed E.coli)<br>
 +
<b>2017-07-17</b>&nbsp;&nbsp;&nbsp; (1) digestion again: BBa_K1356005, BBa_K112806, and BBa_K112000<br>
 +
　　　　　(2) retransform BBa_K112806 and BBa_K112000<br>
 +
<b>2017-07-18</b>&nbsp;&nbsp;&nbsp; (1) proliferate BBa_K112806 transformed E.coli (BBa_K112000 failed)<br>
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　　　　　(2) subculture <i>Synechococcus elongatus</i> PCC7942<br>
 +
　　　　　(3) cultivate <i>Synechocystis</i> sp. PCC6803 in BG11<br>
 +
<b>2017-07-19</b>&nbsp;&nbsp;&nbsp; (1) extract 07-18 BBa_K112806 transformed E.coli plasmid<br>
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　　　　　(2) digestion check<br>
 +
　　　　　(3) save BBa_K112806 transformed E.coli<br>
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　　　　　(4) save 07-18 <i>Synechocystis</i> sp. PCC6803<br>
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<b>2017-07-20</b>&nbsp;&nbsp; (1) activate Rhizobium etli in PY medium<br>
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　　　　　(2) dilute primer 001~008<br>
 +
　　　　　(3) run NrtA and terminator PCR (failed)<br>
 +
<b>2017-07-21 to 07-23</b>&nbsp;&nbsp; cultivate <i>Chlorella vulgaris</i> again for measuring growth curve<br>
 +
<b>2017-07-24</b>&nbsp;&nbsp; (1) prepare <i>Rhizobium etli</i> liquid culture<br>
 +
　　　　　(2) activate <i>Synechococcus elongatus</i> PCC7942<br>
 +
<b>2017-07-25</b>&nbsp;&nbsp; (1) extract <i>Synechocystis</i> sp. PCC6803 genomic DNA<br>
 +
　　　　　(2) run NrtA PCR<br>
 +
<b>2017-07-26</b>&nbsp;&nbsp; (1) extract <i>Rhizobium etli</i> genomic DNA<br>
 +
　　　　　(2) run NrtA and terminator PCR<br>
 +
　　　　　(3) run NrtA+ terminator fusion PCR (failed)<br>
 +
<b>2017-07-27</b>&nbsp;&nbsp; (1) run NrtA+ terminator fusion PCR again<br>
 +
　　　　　(2) purify the successful fusion product <br>
 +
　　　　　(3) digestion check<br>
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<b>2017-07-28</b>&nbsp;&nbsp; run the whole NrtA cloning procedure again, using KOD polymerase<br>
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　　　　　(1) run NrtA and terminator PCR<br>
 +
　　　　　(2) digestion check NrtA and terminator and purify the PCR product<br>
 +
　　　　　(3) run NrtA and terminator fusion PCR<br>
 +
　　　　　(4) digestion check and purify the fusion PCR product (failed)<br>
 +
<b>2017-07-29</b>&nbsp;&nbsp; (1) run NrtA and terminator fusion PCR again<br>
 +
　　　　　(2) digestion check and purify the fusion PCR product<br>
 +
　　　　　(3) ligation: backbone(psb1C3) and promoter, RBS, NrtA, and terminator<br>
 +
　　　　　(4) transformation in to E.coli<br>
 +
<b>2017-07-30</b>&nbsp;&nbsp; cultivate the transformed E.coli in liquid culture<br>
 +
<b>2017-07-31</b>&nbsp;&nbsp;&nbsp; (1) extract 07-30 transformed E.coli plasmid<br>
 +
　　　　　(2) restriction enzyme check (failed)<br>
 +
</p>
 
 
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<div class="panel panel-default">
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<center>
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<div class="panel-heading" role="tab" id="headingOne">
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<a href="#!" onclick="toggleHeight2(this,1440);" style='font-size:20px;color:#2c498c'>
−
<h4 class="panel-title">
+
click to close
−
<a role="button" data-toggle="collapse" data-parent="#accordion" href="#collapseOne" aria-expanded="true" aria-controls="collapseOne">
+
</a>
−
June, 2017
+
</center>
−
</a>
+
</div>
−
</h4>
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</div>
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</div>
+
−
<div id="collapseOne" class="panel-collapse collapse in" role="tabpanel" aria-labelledby="headingOne">
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<div class='panel'>
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<div class="panel-body">
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<div id="s3" class="expandable" style='height: 24px;padding-top:5px;'>
−
What we has done in June...
+
<a href="#!" onclick="toggleHeight3(this, 2200); return false"  
−
Will soon be uploaded...
+
style="font-family: 'Acme', sans-serif;font-size:28px;color: #479fcc;height: 20px;">
−
</div>
+
August, 2017
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</div>
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</a>
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</div>
+
<p>
−
+
<br>
−
<div class="panel panel-default">
+
<b>2017-08-04</b> &nbsp;&nbsp; Digest check: BBa_K112806<br>
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<div class="panel-heading" role="tab" id="headingTwo">
+
<b>2017-08-05</b> &nbsp;&nbsp; <i>Chlorella Vulgaris</i> growth curve measuring<br>
−
<h4 class="panel-title">
+
<b>2017-08-06</b> &nbsp;&nbsp; PCR(Dream Taq): Sucide mechanism-BBa_K112806, K737007, J04450, K112000<br>  
−
<a class="collapsed" role="button" data-toggle="collapse" data-parent="#accordion" href="#collapseTwo" aria-expanded="false" aria-controls="collapseTwo">
+
<b>2017-08-07</b> &nbsp; (1) PCR(KOD plus): Sucide mechanism-BBa_K112806, K737007, J04450, K112000
−
July, 2017
+
<br>　　　　　(2) NrtA- annealing, PNK, ligation, transformation<br>
−
</a>
+
<b>2017-08-08</b> &nbsp;&nbsp; Suicide mechanism-Annealing, cloning, ligation, transformation<br>
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</h4>
+
<b>2017-08-09</b> &nbsp;&nbsp; Extraction of <i>Synechoccocus</i> PCC7942 genomic DNA<br>
−
</div>
+
<b>2017-08-10</b> &nbsp;&nbsp; (1) <i>Chlorella Vulgaris</i> growth curve measuring
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<div id="collapseTwo" class="panel-collapse collapse" role="tabpanel" aria-labelledby="headingTwo">
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<br>　　　　　(2) Fusion PCR: NrtA + terminator
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<div class="panel-body">
+
<br>　　　　　(3) Digestion check: NrtA cut with BamH1
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What we has done in July...
+
<br>　　　　　(4) Subculture <i>S.elongatus</i> PCC7942<br>
−
Will soon be uploaded...
+
<b>2017-08-11</b> &nbsp;&nbsp;&nbsp; (1) Ligation: NrtA + terminator + promoter + RBS
−
</div>
+
<br>　　　　　(2) Ligation: BBa_K737007+K112806, J04450+K112000 (Endolysin and Holin)
−
</div>
+
<br>　　　　　(3) Transformation of Endolysin and Holin
−
</div>
+
<br>　　　　　(4) Measure nitrate and nitrite concentration of <i>Chlorella Vulgaris</i>  
−
+
<br>　　　　　(5) OD value measurement of <i>Chlorella Vulgaris</i> and <i>S.elongatus</i> PCC7942<br>
−
<div class="panel panel-default">
+
<b>2017-08-12</b> &nbsp;&nbsp; (1) Measure nitrate and nitrite concentration of <i>Chlorella Vulgaris</i>  
−
<div class="panel-heading" role="tab" id="headingThree">
+
<br>　　　　　(2) OD value measurement of <i>Chlorella Vulgaris</i> and <i>S.elongatus</i> PCC7942<br>
−
<h4 class="panel-title">
+
<b>2017-08-13</b> &nbsp;&nbsp; (1) Transformation: BBa_K274002 and K1152008 into E. coli
−
<a class="collapsed" role="button" data-toggle="collapse" data-parent="#accordion" href="#collapseThree" aria-expanded="false" aria-controls="collapseThree">
+
<br>　　　　　(2) Measure nitrate and nitrite concentration of <i>Chlorella Vulgaris</i>  
−
Augest, 2017
+
<br>　　　　　(3) OD value measurement of <i>Chlorella Vulgaris</i> and <i>S.elongatus</i> PCC7942<br>
−
</a>
+
<b>2017-08-14</b> &nbsp;&nbsp; (1) Colony PCR: K1152008
−
</h4>
+
<br>　　　　　(2) Suicide mechanism- PCR, annealing, cloning, ligation, transformation
−
</div>
+
<br>　　　　　(3) NrtA-PCR, annealing, cloning, ligation, transformation
−
<div id="collapseThree" class="panel-collapse collapse" role="tabpanel" aria-labelledby="headingThree">
+
<br>　　　　　(4) Liquid culture-K1152008<br>
−
<div class="panel-body">
+
<b>2017-08-15</b> &nbsp;&nbsp; (1) Measure nitrate and nitrite concentration of <i>Chlorella Vulgaris</i>  
−
Will soon be uploaded...
+
<br>　　　　　(2) OD value measurement of <i>Chlorella Vulgaris</i> and <i>S.elongatus</i> PCC7942
−
</div>
+
<br>　　　　　(3) Suicide mechanism- final annealing, cloning, ligation, transformation
−
</div>
+
<br>　　　　　(4) Plasmid extraction and RE check: k1152008<br>
−
</div>
+
<b>2017-08-16</b> &nbsp;&nbsp; (1) Measure nitrate and nitrite concentration of <i>Chlorella Vulgaris</i>  
−
+
<br>　　　　　(2) OD value measurement of Chlorella Vulgaris and <i>S.elongatus</i> PCC7942
−
<div class="panel panel-default">
+
<br>　　　　　(3) PCR(Dream Taq): PrbCL and indC<br>
−
<div class="panel-heading" role="tab" id="headingFour">
+
<b>2017-08-18</b> &nbsp;&nbsp; (1) Suicide mechanism- PCR, annealing, cloning, ligation, transformation
−
<h4 class="panel-title">
+
<br>　　　　　(2) PCR(KOD plus): PrbCL and indC<br>
−
<a class="collapsed" role="button" data-toggle="collapse" data-parent="#accordion" href="#collapseFour" aria-expanded="false" aria-controls="collapseFour">
+
<b>2017-08-19</b> &nbsp;&nbsp; (1) Suicide mechanism- PCR, annealing, cloning, ligation, transformation
−
September, 2017
+
<br>　　　　　(2) Inoculate E. coli containing pBAD18
−
</a>
+
<br>　　　　　(3) Fusion PCR: PrbCL+indC<br>
−
</h4>
+
<b>2017-08-22</b> &nbsp;&nbsp; (1) PCR: Lycopene and MelA
−
</div>
+
<br>　　　　　(2) Fusion PCR: PrbCL+indC<br>
−
<div id="collapseFour" class="panel-collapse collapse" role="tabpanel" aria-labelledby="headingFour">
+
<b>2017-08-24</b> &nbsp;&nbsp; (1) Liquid culture: BBa_K274002
−
<div class="panel-body">
+
<br>　　　　　(2) Extract PCC7942 genomic DNA
−
Will soon be uploaded...
+
<br>　　　　　(3) Inoculate E. coli containing PBR322
−
</div>
+
<br>　　　　　(4) Transformation: BBa_B0015 into E. coli<br>
−
</div>
+
<b>2017-08-25</b> &nbsp;&nbsp; (1) Endolysin- PCR, annealing, cloning, ligation, transformation
−
</div>
+
<br>　　　　　(2) Liquid culture: PBR322
−
+
<br>　　　　　(3) Liquid culture: B0015<br>
−
<div class="panel panel-default">
+
<b>2017-08-26</b> &nbsp;&nbsp; (1) Plasmid extraction: endolysin
−
<div class="panel-heading" role="tab" id="headingFive">
+
<br>　　　　　(2) Plasmid extraction: PBR322
−
<h4 class="panel-title">
+
<br>　　　　　(3) Plasmid extraction: B0015<br>
−
<a class="collapsed" role="button" data-toggle="collapse" data-parent="#accordion" href="#collapseFive" aria-expanded="false" aria-controls="collapseFive">
+
<b>2017-08-27</b> &nbsp;&nbsp; (1) PCR: Lycopene and MelA
−
October, 2017
+
<br>　　　　　(2) Digestion check: PBR322
−
</a>
+
<br>　　　　　(3) Digestion check: B0015
−
</h4>
+
<br>　　　　　(4) pigment backbone primer design
−
</div>
+
<br>　　　　　(5) PCR (Dream Taq): CrtZ
−
<div id="collapseFive" class="panel-collapse collapse" role="tabpanel" aria-labelledby="headingFive">
+
<br>　　　　　(6) Plasmid extraction: endolysin
−
<div class="panel-body">
+
<br>　　　　　(7) Digestion check: endolysin<br>
−
Will soon be uploaded...
+
<b>2017-08-28</b> &nbsp;&nbsp; (1) Colony PCR: NrtA
−
</div>
+
<br>　　　　　(2) Liquid culture: NrtA
−
</div>
+
<br>　　　　　(3) Sequencing: endolysin
−
</div>
+
<br>　　　　　(4) Fusion PCR: PrbcL+IndC
−
+
<br>　　　　　(5) PCR(Dream taq): Holin(kit)
−
</div>
+
<br>　　　　　(6) Check pBR322 concentration and transform pBR322<br>
 +
<b>2017-08-29</b> &nbsp;&nbsp; (1) Nile red oil measuring of <i>Chlorella Vulgaris</i> (~0901)
 +
<br>　　　　　(2) Plasmid extraction: NrtA
 +
<br>　　　　　(3) Holin construction
 +
<br>　　　　　(4) sequencing: NrtA<br>
 +
<b>2017-08-30</b> &nbsp;&nbsp; (1) PCR (Dream Taq): Ampicillin resistance, 5NSII, 3NSII, B0015, and PBR322 ori
 +
<br>　　　　　(2) Endolysin construct finish
 +
<br>　　　　　(3) Plasmid extraction: pBR322
 +
<br>　　　　　(4) Restriction enzyme check: pBR322
 +
<br>　　　　　(5) Backbone construct-3NSII, 5NSII,AMP, B0015, and ORI (PCR: dream taq)<br>
 +
<b>2017-08-31</b> &nbsp;&nbsp; (1) PCR (KOD plus): Ampicillin, 5NSII, 3NSII, B0015, and PBR322 ori
 +
<br>　　　　　(2) PCR (Dream Taq): PBR322 ori and B0015
 +
<br>　　　　　(3) pSB1C3-NrtA construct finish
 +
<br>　　　　　(4) Backbone construct-3NSII, 5NSII, AMP, B0015, and ORI (PCR: KOD)
 +
<br>　　　　　(5) Transform: BBa_K112000
 +
<br>　　　　　(6) LBA: glucose + CM (0.3%)
 +
</p>
 
 
 +
<center>
 +
<a href="#!" onclick="toggleHeight3(this, 2200);" style='font-size:20px;color:#2c498c'>
 +
click to close
 +
</a>
 +
</center>
 +
</div>
 
</div>
 
</div>
 
 
−
<div id="Labnote" class="container-fluid">
+
<div class='panel'>
−
<h1>Labnote</h1>
+
<div id="s4" class="expandable" style='height: 24px;padding-top:5px;'>
−
<a herf="https://static.igem.org/mediawiki/2017/7/7d/T--NYMU-Taipei--Labnote001.pdf">Download the file</a>
+
<a href="#!" onclick="toggleHeight4(this, 1900); return false"
−
<p></p>
+
style="font-family: 'Acme', sans-serif;font-size:28px;color: #479fcc;height: 20px;">
−
<embed src="https://static.igem.org/mediawiki/2017/7/7d/T--NYMU-Taipei--Labnote001.pdf" style="width:90%;height:800px;padding-left:10%">
+
September, 2017
 +
</a>
 +
<p>
 +
<br>
 +
<b>2017-09-01</b> &nbsp;&nbsp; (1)Fusion PCR: Ampicillin resistance, B0015
 +
<br>　　　　　(2) Three pieces PCR: 5NSII, 3NSII, and PBR322 ori<br>
 +
 
 +
<b>2017-09-02</b> &nbsp; (1) Ligation: Ampicillin resistance, B0015+ 5NSII, 3NSII, and PBR322 ori
 +
<br>　　　　　(2) Transformation: pPIGBACK (pigment plasmid backbone)
 +
<br>　　　　　(3) PCR (Dream Taq): CrtZ and PrbCL<br>
 +
 
 +
<b>2017-09-03</b> &nbsp; (1) Colony PCR: pPIGBACK
 +
<br>　　　　　(2) Liquid culture: pPIGBACK
 +
<br>　　　　　(3) PCR (KOD): CrtZ and PrbCL<br>
 +
 
 +
<b>2017-09-04</b> &nbsp; (1) Plasmid extraction: pPIGBACK
 +
<br>　　　　　(2) Digestion check: pPIGBACK
 +
<br>　　　　　(3) Fusion PCR: CrtZ+ PrbCL<br>
 +
 
 +
<b>2017-09-05</b> &nbsp; (1) pPIGBACK construct finish
 +
<br>　　　　　(2) Fusion PCR: PrbCL+IndC
 +
<br>　　　　　(3) Ligation: CrtZ+ PrbCL with pPIGBACK
 +
<br>　　　　　(4) Transformation: CrtZ construct<br>
 +
 
 +
<b>2017-09-06</b> &nbsp; (1) Colony pcr: CrtZ construct
 +
<br>　　　　　(2) Liquid culture: CrtZ
 +
<br>　　　　　(3) Lycopene primers design
 +
<br>　　　　　(4) IndC primers redesign<br>
 +
 
 +
<b>2017-09-07</b> &nbsp; (1) plasmid extraction: CrtZ-Ppigback
 +
<br>　　　　　(2) restriction enzyme check: CrtZ-pPIGBACK<br>
 +
 
 +
<b>2017-09-08</b> &nbsp; (1) transform CrtZ-Ppigback into PCC7942
 +
<br>　　　　　(2) nitrate and nitrite adsorption assay of Chlorella vulgaris (co-coculture with NrtA)
 +
<br>　　　　　(3) transform suicide mechanism construct (LBA+glucose 0.3%)<br>  
 +
 
 +
<b>2017-09-09</b> &nbsp; (1) nitrate and nitrite adsorption assay of Chlorella vulgaris (co-coculture with NrtA)
 +
<br>　　　　　(2) fusion PCR: PrbcL+indC<br>
 +
 
 +
<b>2017-09-10</b> &nbsp;&nbsp; indC construct (~transformation)<br>
 +
 
 +
<b>2017-09-19</b> &nbsp;&nbsp; (1) Endolysin-Holin construct
 +
<br>　　　　　(2) Site-directed primers design<br>
 +
 
 +
<b>2017-09-20</b> &nbsp; (1) Liquid culture: BBa_J04450
 +
<br>　　　　　(2) Liquid culture: CrtZ-PCC7942 clones<br>
 +
 
 +
<b>2017-09-21</b> &nbsp;&nbsp; Plasmid extraction: J04450, CrtZ-PCC7942<br>
 +
 
 +
<b>2017-09-22</b> &nbsp;&nbsp; Site-directed PCR: 3NSII<br>
 +
 
 +
<b>2017-09-23</b> &nbsp;&nbsp; Site-directed-3NSII construct finish<br>
 +
 
 +
<b>2017-09-24</b> &nbsp; (1) plasmid extraction: Site-directed-3NSII
 +
<br>　　　　　(2) Site-directed PCR: 5NSII-1
 +
<br>　　　　　(3) Psb1c3-PrbcL construct finish<br>
 +
 
 +
<b>2017-09-25</b> &nbsp; (1) Psb1c3-NrtA construct finish
 +
<br>　　　　　(2) colony PCR: Psb1c3-PrbcL
 +
<br>　　　　　(3) Liquid culture: Psb1c3-PrbcL
 +
<br>　　　　　(4) IndC construct<br>
 +
 
 +
<b>2017-09-26</b> &nbsp; (1) Plasmid extraction and restriction enzyme check: Site-directed-5NSII-1
 +
<br>　　　　　(2) Liquid culture: Psb1c3-NrtA
 +
<br>　　　　　(3) Colony PCR: IndC
 +
<br>　　　　　(4) Psb1c3-3NSII construct finish
 +
<br>　　　　　(5) Site-directed PCR: 5NSII-2<br>
 +
 
 +
<b>2017-09-27</b> &nbsp; (1) Psb1c3-NrtA construct finish
 +
<br>　　　　　(2) Plasmid extraction and restriction enzyme check: IndC
 +
<br>　　　　　(3) Plasmid extraction and restriction enzyme check: IndC
 +
<br>　　　　　(4) Psb1c3-3NSII construct finish Plasmid extraction and RE check: Site-directed 5NSII-2
 +
<br>　　　　　(5) Site-directed PCR: AmpR<br>
 +
 
 +
<b>2017-09-28</b> &nbsp; (1) Plasmid extraction and RE check: Site-directed-AmpR
 +
<br>　　　　　(2) Plasmid extraction and RE check: Psb1c3-5NSII
 +
<br>　　　　　(3) Psb1c3-3NSII(SD) finish
 +
<br>　　　　　(4) Liquid culture: Psb1c3-PrbcL<br>
 +
 
 +
<b>2017-09-29</b> &nbsp; (1) Psb1c3-5NSII construct finish
 +
<br>　　　　　(2) Psb1c3-5NSII-ORI-3NSII construct finish
 +
<br>　　　　　(3) Colony PCR: Psb1c3-PrbcL- IndC
 +
<br>　　　　　(4) Colony PCR: Psb1c3-PrbcL- IndC
 +
<br>　　　　　(5) Psb1c3-PrbcL(SD) construct finish
 +
<br>　　　　　(6) Psb1c3-PrbcL-IndC construct finish
 +
<br>　　　　　(7) Site-directed: amp finish
 +
<br>　　　　　(8) transform CrtZ-pPIGBACK into PCC7942<br>
 +
 
 +
<b>2017-09-30</b> &nbsp; (1) Psb1c3-PrbcL(SD) finish
 +
<br>　　　　　(2) Liquid culture: CrtZ-pPIGBACK
 +
 
 +
</p>
 +
 +
<center>
 +
<a href="#!" onclick="toggleHeight4(this, 1900);" style='font-size:20px;color:#2c498c'>
 +
click to close
 +
</a>
 +
</center>
 
</div>
 
</div>
−
+
</div>
−
+
 +
<div class='panel'>
 +
<div id="s5" class="expandable" style='height: 24px;padding-top:5px;'>
 +
<a href="#!" onclick="toggleHeight5(this, 2240); return false"
 +
style="font-family: 'Acme', sans-serif;font-size:28px;color: #479fcc;height: 20px;">
 +
October, 2017
 +
</a>
 +
<p>
 +
<br>
 +
<b>2017-10-01</b> &nbsp;&nbsp; (1) Colony PCR: 5NSII-Psb1c3
 +
<br>　　　　　(2) Liquid culture: 5NSII-Psb1c3
 +
<br>　　　　　(3) Colony PCR: 5NSII-ORI-3NSII-Psb1c3
 +
<br>　　　　　(4) Liquid culture: Site-directed mutated clones
 +
<br>　　　　　(5) Construct: Psb1c3-PrbcL-IndC
 +
<br>　　　　　(6) Liquid culture: NrtA-C33
 +
 
 +
<br><b>2017-10-02</b> &nbsp;&nbsp; (1) Finish all aite-directed mutagenesis
 +
<br>　　　　　(2) plasmid extraction: Psb1c3-5NSII, 5NSII-ORI-3NSII-Psb1c3
 +
<br>　　　　　(3) Digestion check: Psb1c3-5NSII, 5NSII-ORI-3NSII-Psb1c3
 +
<br>　　　　　(4) Colony PCR: Psb1c3-PrbcL-IndC
 +
<br>　　　　　(5) Liquid culture: Psb1c3-PrbcL-IndC
 +
<br>　　　　　(6) Liquid culture (French Press): NrtA-C33, competent cell
 +
 
 +
<br><b>2017-10-03</b> &nbsp;&nbsp; (1) French Press and dialysis
 +
<br>　　　　　(2) Sequencing: MelA
 +
<br>　　　　　(3) Construct Psb1c3-NrtA-Endolysin
 +
 
 +
<br><b>2017-10-04</b> &nbsp;&nbsp; (1) Construct pPIGBACK-J04450
 +
<br>　　　　　(2) Construct Psb1c3-B0015-AMP
 +
<br>　　　　　(3) Construct Psb1c3-PrbcL-CrtZ
 +
<br>　　　　　(4) check nitrate concentration
 +
<br>　　　　　(5) Colony PCR: NrtA-endolysin
 +
<br>　　　　　(6) Liquid culture: NrtA-endolysin
 +
 
 +
<br><b>2017-10-05</b> &nbsp;&nbsp; (1) Colony PCR: pPIGBACK-J04450
 +
<br>　　　　　(2) Liquid culture: pPIGBACK-J04450
 +
<br>　　　　　(3) Digestion check: NrtA-endolysin
 +
 
 +
<br><b>2017-10-06</b> &nbsp;&nbsp; (1) DNA extraction and digestion-pPIGBACK-CrtZ
 +
<br>　　　　　(2) Finish construct pPIGBACK-J04450
 +
<br>　　　　　(3) Colony PCR: Psb1c3-B0015-AMP
 +
<br>　　　　　(4) Liquid culture: Psb1c3-B0015-AMP
 +
 
 +
<br><b>2017-10-07</b> &nbsp;&nbsp; (1) Plasmid extraction: Psb1c3-B0015-AMP
 +
<br>　　　　　(2) Digest check: Psb1c3-B0015-AMP
 +
<br>　　　　　(3) Construct: Psb1c3-PrbcL-CrtZ
 +
 
 +
<br><b>2017-10-08</b> &nbsp;&nbsp; (1) Colony PCR: Psb1c3-PrbcL-CrtZ
 +
 
 +
<br><b>2017-10-09</b> &nbsp;&nbsp; (1) Liquid culture: competent cell for electroporation
 +
<br>　　　　　(2) Frech Press
 +
<br>　　　　　(3) Nitrate concentration assay
 +
 
 +
<br><b>2017-10-10</b> &nbsp;&nbsp; (1) Electroporation: competent cell
 +
 
 +
<br><b>2017-10-11</b> &nbsp;&nbsp; (1) Test electroporation: Psb1c3-J04450
 +
 
 +
<br><b>2017-10-13</b> &nbsp;&nbsp; (1) Construct: Psb1c3-IndC (electroporation)
 +
<br>　　　　　(2) Construct: pPIGBACK-IndC (electroporation)
 +
<br>　　　　　(3) Construct: pPIGBACK-PrbcL-CrtZ
 +
<br>　　　　　(4) Construct: Psb1c3-B0015-AMP
 +
<br>　　　　　(5) Starch functional test: pPIGBACK-PrbcL-CrtZ
 +
<br>　　　　　(6) Suicide mechanism functional test
 +
 
 +
<br><b>2017-10-14</b> &nbsp;&nbsp; (1) Colony PCR: pPIGBACK-IndC
 +
<br>　　　　　(2) Liquid culture: pPIGBACK-IndC
 +
<br>　　　　　(3) Colony PCR: pPIGBACK-PrbcL-CrtZ
 +
<br>　　　　　(4) <i>Chlorella Vulgaris</i> oil functional test: NrtA
 +
<br>　　　　　(5) Starch functional test: pPIGBACK-PrbcL-CrtZ
 +
<br>　　　　　(6) Suicide mechanism functional test
 +
 
 +
<br><b>2017-10-15</b> &nbsp;&nbsp; (1) Finish construct pPIGBACK-PrbcL-IndC, Psb1c3-PrbcL-IndC
 +
<br>　　　　　(2) Liquid culture: All parts
 +
<br>　　　　　(3) Starch functional test: pPIGBACK-PrbcL-CrtZ
 +
 
 +
<br><b>2017-10-16</b> &nbsp;&nbsp; (1) <i>Chlorella Vulgaris</i> oil functional test: NrtA
 +
<br>　　　　　(2) Plasmid extraction: all parts
 +
<br>　　　　　(3) Digestion check: all parts
 +
<br>　　　　　(4) Starch functional test: pPIGBACK-PrbcL-CrtZ
 +
 
 +
<br><b>2017-10-17</b> &nbsp;&nbsp; (1) NrtA add to device again
 +
<br>　　　　　(2) Starch functional test: pPIGBACK-PrbcL-CrtZ
 +
 
 +
<br><b>2017-10-18</b> &nbsp;&nbsp; (1) NrtA add to device again
 +
<br>　　　　　(2) Starch functional test: pPIGBACK-PrbcL-CrtZ
 +
 
 +
<br><b>2017-10-19</b> &nbsp;&nbsp; (1) NrtA add to device again
 +
<br>　　　　　(2) Starch functional test: pPIGBACK-PrbcL-CrtZ
 +
 
 +
<br><b>2017-10-20</b> &nbsp;&nbsp; (1) NrtA add to device again
 +
<br>　　　　　(2) Starch functional test: pPIGBACK-PrbcL-CrtZ
 +
 
 +
<br><b>2017-10-21</b> &nbsp;&nbsp; (1) NrtA add to device again
 +
<br>　　　　　(2) Starch functional test: pPIGBACK-PrbcL-CrtZ
 +
 
 +
<br><b>2017-10-22</b> &nbsp;&nbsp; (1) PCR: MelA, IndC
 +
<br>　　　　　(2) Ligation: Psb1c3-MelA, pPIGBACK-MelA, Psb1c3-IndC, pPIGBACK-IndC
 +
<br>　　　　　(3) Transformation: Psb1c3-MelA, pPIGBACK-MelA, Psb1c3-IndC, pPIGBACK-IndC
 +
 
 +
<br><b>2017-10-23</b> &nbsp;&nbsp; (1) PCR: MelA, IndC, Psb1c3-B0015-AMP
 +
<br>　　　　　(2) Ligation: Psb1c3-MelA, pPIGBACK-MelA, Psb1c3-IndC, pPIGBACK-IndC, Psb1c3-B0015-AMP
 +
<br>　　　　　(3) Liquid culture: Psb1c3-MelA, pPIGBACK-MelA, Psb1c3-IndC, pPIGBACK-IndC
 +
 
 +
<br><b>2017-10-24</b> &nbsp;&nbsp; (1) PCR: PrbcL-MelA, Psb1c3-IndC, pPIGBACK-IndC
 +
<br>　　　　　(2) Ligation: Psb1c3-PrbcL-MelA, pPIGBACK-PrbcL-MelA
 +
<br>　　　　　(3) Transformation: Psb1c3-PrbcL-MelA, pPIGBACK-PrbcL-MelA, Psb1c3-IndC, pPIGBACK-IndC, Psb1c3-B0015-AMP
 +
<br>　　　　　(4) Liquid culture: Psb1c3-B0015-AMP
 +
 
 +
<br><b>2017-10-25</b> &nbsp;&nbsp; (1) Digestion check all parts
 +
<br>　　　　　(2) Plasmid extraction: Psb1c3-B0015-AMP
 +
<br>　　　　　(3) Digestion check: Psb1c3-B0015-AMP
 +
 
 +
</p>
 +
 +
<center>
 +
<a href="#!" onclick="toggleHeight5(this, 2240);" style='font-size:20px;color:#2c498c'>
 +
click to close
 +
</a>
 +
</center>
 +
</div>
 +
</div>
 +
 +
</div>
 
 
 +
<div id="Labnote">
 +
<h1>Labnote</h1>
 +
<p></p>
 +
<a href="https://static.igem.org/mediawiki/2017/0/04/T--NYMU-Taipei--labnote.pdf" target="_blank">Download the file</a>
 +
<p></p>
 +
<embed src="https://static.igem.org/mediawiki/2017/0/04/T--NYMU-Taipei--labnote.pdf" style="width:80%;height:500px;padding-left:10%">
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Latest revision as of 16:14, 28 November 2017

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