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<meta name="renderer" content="webkit"> | <meta name="renderer" content="webkit"> | ||
<title>Interlab</title> | <title>Interlab</title> | ||
− | + | <link rel="stylesheet" href="https://2017.igem.org/Team:HUST-China/css/bootstrap?action=raw&ctype=text/css"> | |
− | + | <link href="https://2017.igem.org/Team:HUST-China/css/return_top?action=raw&ctype=text/css" rel="stylesheet" type="text/css" /> | |
− | + | <link rel="stylesheet" href="https://2017.igem.org/Team:HUST-China/css/picbox.css?action=raw&ctype=text/css" type="text/css" media="screen" /> | |
− | < | + | |
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<script> | <script> | ||
if ($('.firstHeading')) $('.firstHeading').remove(); | if ($('.firstHeading')) $('.firstHeading').remove(); | ||
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padding: 20px; | padding: 20px; | ||
text-align: center; | text-align: center; | ||
+ | } | ||
+ | .text h4{ | ||
+ | padding: 5px 10px; | ||
+ | } | ||
+ | .text p{ | ||
+ | padding: 2px 10px; | ||
+ | } | ||
+ | .text h5{ | ||
+ | padding: 3px 10px; | ||
+ | } | ||
+ | .dropdown button{ | ||
+ | margin: 2px 20px; | ||
} | } | ||
/*没有小于768px,是因为Bootstrap3以移动端优先设计*/ | /*没有小于768px,是因为Bootstrap3以移动端优先设计*/ | ||
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} | } | ||
.text p { | .text p { | ||
− | font-size: | + | font-size: 10px; |
− | text-indent: | + | text-indent: 20px; |
} | } | ||
.tab2-text { | .tab2-text { | ||
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} | } | ||
.text p { | .text p { | ||
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.text p { | .text p { | ||
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} | } | ||
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position: fixed; | position: fixed; | ||
} | } | ||
− | div.col-sm- | + | ul.nav-pills2 { |
+ | bottom:50px; | ||
+ | position: fixed; | ||
+ | } | ||
+ | div.col-sm-10 div { | ||
min-height: 100px; | min-height: 100px; | ||
font-size: 14px; | font-size: 14px; | ||
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} | } | ||
− | @media screen and (max-width: | + | @media screen and (max-width: 992px) { |
#section1, #section2, #section3, #section4, #section5 { | #section1, #section2, #section3, #section4, #section5 { | ||
margin-left: 50px; background-color: #eee; color:#171717; | margin-left: 50px; background-color: #eee; color:#171717; | ||
} | } | ||
+ | } | ||
+ | .yuandian li {list-style-type:circle; } | ||
+ | |||
+ | body { | ||
+ | position: relative; | ||
+ | } | ||
+ | @media (min-width: 768px) { | ||
+ | .menu_logo_1 { | ||
+ | top: 18px; | ||
+ | left:10px; | ||
+ | z-index: 1000; | ||
+ | position: fixed; | ||
+ | } | ||
+ | .menu_logo_2 { | ||
+ | top: 18px; | ||
+ | left:100px; | ||
+ | z-index: 1000; | ||
+ | position: fixed; | ||
+ | } | ||
+ | .nav-pills{ | ||
+ | font-size:8px; | ||
+ | font-weight: bold; | ||
+ | } | ||
+ | } | ||
+ | |||
+ | /* 中等屏幕(桌面显示器,大于等于992px) */ | ||
+ | @media (min-width: 992px) { | ||
+ | .menu_logo_1 { | ||
+ | top: 18px; | ||
+ | left:20px; | ||
+ | z-index: 1000; | ||
+ | position: fixed; | ||
+ | } | ||
+ | .menu_logo_2 { | ||
+ | top: 18px; | ||
+ | left:110px; | ||
+ | z-index: 1000; | ||
+ | position: fixed; | ||
+ | } | ||
+ | .nav-pills{ | ||
+ | font-size:10px; | ||
+ | font-weight: bold; | ||
+ | } | ||
+ | } | ||
+ | |||
+ | /* 大屏幕(大桌面显示器,大于等于1200px) */ | ||
+ | @media (min-width: 1200px) { | ||
+ | .menu_logo_1 { | ||
+ | top: 18px; | ||
+ | left:80px; | ||
+ | z-index: 1000; | ||
+ | position: fixed; | ||
+ | } | ||
+ | .menu_logo_2 { | ||
+ | top: 18px; | ||
+ | left:170px; | ||
+ | z-index: 1000; | ||
+ | position: fixed; | ||
+ | } | ||
+ | .nav-pills{ | ||
+ | font-size:12px; | ||
+ | font-weight: bold; | ||
+ | } | ||
} | } | ||
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<body data-spy="scroll" data-target="#myScrollspy" data-offset="20"> | <body data-spy="scroll" data-target="#myScrollspy" data-offset="20"> | ||
− | + | <nav class="navbar navbar-default navbar-fixed-top" style="margin-top:15px"> | |
<div class="container"> | <div class="container"> | ||
− | |||
<div class="navbar-header"> | <div class="navbar-header"> | ||
− | <a href="https://2017.igem.org/Team:HUST-China" class="navbar-brand logo"><img src="https://static.igem.org/mediawiki/2017/6/64/Team_HUST_China_2017_logo.png" alt="HUST_iGEM_2017_logo"></a> | + | <a href="https://2017.igem.org/Team:HUST-China" class="navbar-brand logo menu_logo_1"><img src="https://static.igem.org/mediawiki/2017/6/64/Team_HUST_China_2017_logo.png" alt="HUST_iGEM_2017_logo"></a> |
− | <button type="button" class="navbar-toggle" data-toggle="collapse" data-target="#navbar-collapse"> | + | <a href="https://2017.igem.org" class="navbar-brand logo menu_logo_2"><img src="https://static.igem.org/mediawiki/2017/f/f6/IGEM_2017_logo.png" alt="HUST_iGEM_2017_logo"></a> |
− | <span class=" | + | <button type="button" class="navbar-toggle" data-toggle="collapse" data-target="#navbar-collapse" style="background-color: #333; color:#111"> |
+ | <span class="icon-bar"></span> | ||
+ | <span class="icon-bar"></span> | ||
+ | <span class="icon-bar"></span> | ||
+ | <span class="icon-bar"></span> | ||
</button> | </button> | ||
</div> | </div> | ||
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<div class="collapse navbar-collapse" id="navbar-collapse"> | <div class="collapse navbar-collapse" id="navbar-collapse"> | ||
<ul class="nav navbar-nav navbar-right"> | <ul class="nav navbar-nav navbar-right"> | ||
+ | <li><a href="https://2017.igem.org/Team:HUST-China">HOME</a></li> | ||
+ | <li class="dropdown active"> | ||
+ | <a href="#" class="dropdown-toggle" data-toggle="dropdown"> | ||
+ | PROJECT <b class="caret"></b> | ||
+ | </a> | ||
+ | <ul class="dropdown-menu"> | ||
+ | <li><a href="https://2017.igem.org/Team:HUST-China/project/background">Background</a></li> | ||
+ | <li><a href="https://2017.igem.org/Team:HUST-China/Description">Description</a></li> | ||
+ | <li><a href="https://2017.igem.org/Team:HUST-China/Safety">Safety</a></li> | ||
+ | <li><a href="https://2017.igem.org/Team:HUST-China/Demonstrate">Demonstrate</a></li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | <li class="dropdown"> | ||
+ | <a href="#" class="dropdown-toggle" data-toggle="dropdown"> | ||
+ | WETLAB <b class="caret"></b> | ||
+ | </a> | ||
+ | <ul class<nav class="navbar navbar-default navbar-fixed-top" style="margin-top:15px"> | ||
+ | <div class="container"> | ||
+ | <div class="navbar-header"> | ||
+ | <a href="https://2017.igem.org/Team:HUST-China" class="navbar-brand logo menu_logo_1"><img src="https://static.igem.org/mediawiki/2017/6/64/Team_HUST_China_2017_logo.png" alt="HUST_iGEM_2017_logo"></a> | ||
+ | <a href="https://2017.igem.org" class="navbar-brand logo menu_logo_2"><img src="https://static.igem.org/mediawiki/2017/f/f6/IGEM_2017_logo.png" alt="HUST_iGEM_2017_logo"></a> | ||
+ | <button type="button" class="navbar-toggle" data-toggle="collapse" data-target="#navbar-collapse" style="background-color: #333; color:#111"> | ||
+ | <span class="icon-bar"></span> | ||
+ | <span class="icon-bar"></span> | ||
+ | <span class="icon-bar"></span> | ||
+ | <span class="icon-bar"></span> | ||
+ | </button> | ||
+ | </div> | ||
− | + | <div class="collapse navbar-collapse" id="navbar-collapse"> | |
+ | <ul class="nav navbar-nav navbar-right"> | ||
+ | <li><a href="https://2017.igem.org/Team:HUST-China">HOME</a></li> | ||
<li class="dropdown"> | <li class="dropdown"> | ||
<a href="#" class="dropdown-toggle" data-toggle="dropdown"> | <a href="#" class="dropdown-toggle" data-toggle="dropdown"> | ||
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<ul class="dropdown-menu"> | <ul class="dropdown-menu"> | ||
<li><a href="https://2017.igem.org/Team:HUST-China/project/background">Background</a></li> | <li><a href="https://2017.igem.org/Team:HUST-China/project/background">Background</a></li> | ||
− | <li><a href="https://2017.igem.org/Team:HUST-China/Description">Description</a></li> | + | <li><a href="https://2017.igem.org/Team:HUST-China/Description">Description</a></li> |
− | + | ||
− | + | ||
− | + | ||
<li><a href="https://2017.igem.org/Team:HUST-China/Safety">Safety</a></li> | <li><a href="https://2017.igem.org/Team:HUST-China/Safety">Safety</a></li> | ||
+ | <li><a href="https://2017.igem.org/Team:HUST-China/Demonstrate">Demonstrate</a></li> | ||
</ul> | </ul> | ||
</li> | </li> | ||
− | <li class="dropdown"> | + | <li class="dropdown active"> |
<a href="#" class="dropdown-toggle" data-toggle="dropdown"> | <a href="#" class="dropdown-toggle" data-toggle="dropdown"> | ||
WETLAB <b class="caret"></b> | WETLAB <b class="caret"></b> | ||
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</li> | </li> | ||
<li class="dropdown"> | <li class="dropdown"> | ||
− | <a href=" | + | <a href="https://2017.igem.org/Team:HUST-China/Model" > |
− | + | MODEL | |
</a> | </a> | ||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
</li> | </li> | ||
<li class="dropdown"> | <li class="dropdown"> | ||
− | + | <a href="#" class="dropdown-toggle" data-toggle="dropdown"> | |
− | PARTS <b class="caret"></b> | + | PARTS<b class="caret"></b> |
</a> | </a> | ||
<ul class="dropdown-menu"> | <ul class="dropdown-menu"> | ||
− | + | <li><a href="https://2017.igem.org/Team:HUST-China/Basic part">Basic part</a></li> | |
− | + | <li><a href="https://2017.igem.org/Team:HUST-China/Composite part">Composite part</a></li> | |
− | + | <li><a href="https://2017.igem.org/Team:HUST-China/Part collection">Part collection</a></li> | |
− | + | ||
</ul> | </ul> | ||
</li> | </li> | ||
<li class="dropdown"> | <li class="dropdown"> | ||
<a href="#" class="dropdown-toggle" data-toggle="dropdown"> | <a href="#" class="dropdown-toggle" data-toggle="dropdown"> | ||
− | + | HP<b class="caret"></b> | |
</a> | </a> | ||
<ul class="dropdown-menu"> | <ul class="dropdown-menu"> | ||
− | + | <li><a href="https://2017.igem.org/Team:HUST-China/Collaborations"> Collaborations</a></li> | |
− | <li><a href="https://2017.igem.org/Team:HUST-China/ | + | <li><a href="https://2017.igem.org/Team:HUST-China/HP/Silver">HP/Silver</a></li> |
+ | <li><a href="https://2017.igem.org/Team:HUST-China/Engagement">Engagement</a></li> | ||
</ul> | </ul> | ||
</li> | </li> | ||
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</nav> | </nav> | ||
− | + | <div class="tab1" id="section0" style="width:100%;margin-top: 65px;"> | |
− | <div class="tab1" style="width:100%;margin-top: 65px;"> | + | |
<h1 class="tab-h1" ><strong>「Interlab」</strong></h1> | <h1 class="tab-h1" ><strong>「Interlab」</strong></h1> | ||
</div> | </div> | ||
− | <div class="container-fluid"> | + | <div class="container-fluid text"> |
<div class="row"> | <div class="row"> | ||
− | <nav class="col-sm- | + | <nav class="col-sm-2" id="myScrollspy"> |
− | + | ||
<div class="container-fluid"> | <div class="container-fluid"> | ||
− | <ul class="nav nav-pills nav-stacked" | + | <div class="collapse navbar-collapse" id="navbar-collapse"> |
+ | <ul class="nav nav-pills nav-stacked" style="font-size:12px; text-align:center; background:#ccc;"> | ||
<li class="active"><a href="#section1">Introduction</a></li> | <li class="active"><a href="#section1">Introduction</a></li> | ||
<li><a href="#section2">Provenance and Release</a></li> | <li><a href="#section2">Provenance and Release</a></li> | ||
− | <li><a href="#section3">Chassis and Safety | + | <li><a href="#section3">Chassis and Safety </a></li> |
− | <li><a href="#section4">Instrument | + | <li><a href="#section4">Instrument</a></li> |
<li><a href="#section5">Calibration Protocol</a></li> | <li><a href="#section5">Calibration Protocol</a></li> | ||
+ | <li><a href="#section6">Cell Culture</a></li> | ||
+ | <li><a href="#section7">Interlab Results</a></li> | ||
</ul> | </ul> | ||
</div> | </div> | ||
</div> | </div> | ||
</nav> | </nav> | ||
− | <div class="col-sm- | + | <div class="col-sm-8"> |
<div id="section1" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | <div id="section1" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | ||
− | <h3 style="background-color:# | + | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px;margin-top:0;">Introduction</h3> |
<p style="padding:10px">It is significant for synthetic biology to develop a reliable and repeatable measurement, the same to all the other engineering disciplines. We HUST-China have volunteered to test some RBS devices (BCDs) that are intended to make gene expression more precise and reliable by measure the expression level of GFP, in order to help the iGEM community collect data about how reliable will these devices turn out to be in labs around the world. | <p style="padding:10px">It is significant for synthetic biology to develop a reliable and repeatable measurement, the same to all the other engineering disciplines. We HUST-China have volunteered to test some RBS devices (BCDs) that are intended to make gene expression more precise and reliable by measure the expression level of GFP, in order to help the iGEM community collect data about how reliable will these devices turn out to be in labs around the world. | ||
</p> | </p> | ||
</div> | </div> | ||
<div id="section2" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | <div id="section2" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | ||
− | <h3 style="background-color:# | + | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Provenance and Release</h3> |
<h4><strong>①Individuals responsible for conducting InterLab study</strong> </h4> | <h4><strong>①Individuals responsible for conducting InterLab study</strong> </h4> | ||
− | <div class="table-responsive" style="padding: 10px 100px; text-align: center;"> | + | <div class="table-responsive" style="padding: 10px 100px; text-align: center;font-size:14px;"> |
− | <table class="table"> | + | <table class="table table-hover"> |
<thead> | <thead> | ||
<tr> | <tr> | ||
− | <th>Individuals</th> | + | <th style="text-align: center;">Individuals</th> |
− | <th>Interlab Part</th> | + | <th style="text-align: center;">Interlab Part</th> |
</tr> | </tr> | ||
</thead> | </thead> | ||
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</div> | </div> | ||
<h4><strong> ②Corresponding email</strong> </h4> | <h4><strong> ②Corresponding email</strong> </h4> | ||
− | <div class="table-responsive" style="padding: 10px 100px; text-align: center;"> | + | <div class="table-responsive" style="padding: 10px 100px; text-align: center;font-size:14px;"> |
− | <table class="table"> | + | <table class="table table-hover"> |
<thead> | <thead> | ||
<tr> | <tr> | ||
− | <th> | + | <th style="text-align: center;">Individuals </th> |
− | <th> | + | <th style="text-align: center;">Emails</th> |
</tr> | </tr> | ||
</thead> | </thead> | ||
<tbody> | <tbody> | ||
+ | <tr> | ||
+ | <td>Efan Wang</td> | ||
+ | <td>erfan@hust.edu.cn</td> | ||
+ | </tr> | ||
<tr> | <tr> | ||
<td>Kangyuan Yu</td> | <td>Kangyuan Yu</td> | ||
− | <td> | + | <td>u201612160@hust.edu.cn</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Haibo Huang </td> | <td>Haibo Huang </td> | ||
− | <td> | + | <td>u201612177@hust.edu.cn</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Ziyang Xiao</td> | <td>Ziyang Xiao</td> | ||
− | <td> | + | <td>u201612166@hust.edu.cn</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Shaofeng Liao</td> | <td>Shaofeng Liao</td> | ||
− | <td> | + | <td>u201612159@hust.edu.cn</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>HuiPing Shi</td> | <td>HuiPing Shi</td> | ||
− | <td> | + | <td>u201513154@hust.edu.cn</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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</div> | </div> | ||
<div id="section3" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | <div id="section3" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | ||
− | <h3 style="background-color:# | + | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Chassis and Safety</h3> |
<h4><strong> What chassis did you use?</strong> </h4> | <h4><strong> What chassis did you use?</strong> </h4> | ||
− | <p>Escherichia coli | + | <p>Escherichia coli DH5α</p> |
<h4><strong> What Biosafety Level is your chassis? </strong> </h4> | <h4><strong> What Biosafety Level is your chassis? </strong> </h4> | ||
<p>BSL1</p> | <p>BSL1</p> | ||
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</div> | </div> | ||
<div id="section4" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | <div id="section4" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | ||
− | <h3 style="background-color:# | + | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Instrument</h3> |
− | <h4><strong> | + | <h4><strong> What instrument did you use during your measurements? </strong> </h4> |
<p>plate reader</p> | <p>plate reader</p> | ||
<h4><strong> Please provide the brand and model of your instrument.</strong> </h4> | <h4><strong> Please provide the brand and model of your instrument.</strong> </h4> | ||
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</div> | </div> | ||
<div id="section5" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | <div id="section5" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | ||
− | <h3 style="background-color:# | + | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Calibration Protocol</h3> |
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− | <h4><strong> A2. Measurement Steps</strong> | + | <h4><strong> A2. Measurement Steps</strong></h4> |
− | + | <li class="dropdown" style="list-style-type:none;"> | |
− | + | <button href="#" class="dropdown-toggle" data-toggle="dropdown"> | |
− | + | More details <b class="caret"></b> | |
− | + | </button> | |
− | + | <ul class="dropdown-menu container" style="padding: 10px;background:#ccc;"> | |
− | + | <li>① Prepare your 96 well plate</li> | |
− | + | <li>②Add 100 μl LUDOX 100 % into wells A1, B1, C1, D1</li> | |
− | + | <li>③Add 100 μl of dH2O into A2, B2, C2, D2</li> | |
− | + | <li>④Measure absorbance 600 nm of all samples in all standard measurement modes in instrument</li> | |
− | + | </ul> | |
− | + | </li> | |
− | + | ||
− | + | ||
<h4><strong> B1. Protocol for FIuorescein Fluoresence standard curve</strong> </h4> | <h4><strong> B1. Protocol for FIuorescein Fluoresence standard curve</strong> </h4> | ||
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<p>22℃</p> | <p>22℃</p> | ||
− | <h4><strong> | + | <h4><strong> B2. Measurement Steps</strong></h4> |
<h5><strong> Part 1: Prepare the Fluorescein stock solution </strong> </h5> | <h5><strong> Part 1: Prepare the Fluorescein stock solution </strong> </h5> | ||
− | + | <li class="dropdown" style="list-style-type:none;"> | |
− | + | <button href="#" class="dropdown-toggle" data-toggle="dropdown"> | |
− | + | More details <b class="caret"></b> | |
− | + | </button> | |
− | + | <ul class="dropdown-menu" style="padding: 10px; background:#ccc;"> | |
− | + | <li>① Spin down fluorescein stock tube to make sure pellet is at the bottom of tube.</li> | |
− | + | <li>② Prepare 2x fluorescein stock solution (100 μM) by resuspending fluorescein in 1ml of 1xPBS</li> | |
− | + | <li>③ Dilute the 2x fluorescein stock solution using 1xPBS to make a 1x fluorescein solution (final concentration is 50 μM).</li> | |
− | + | <li>④ *Illustration of serial dilution samples in 96 well plate or cuvettes: value decreases by 2-fold with each column (50% in column 2, 25% in column 3, 12.5% in column 4, etc.)</li> | |
− | <li> | + | </ul> |
+ | </li> | ||
+ | <h5>Measurement work flow:</h5> | ||
+ | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/> | ||
+ | <a href="https://static.igem.org/mediawiki/2017/f/f8/2017_HUST_China_interlab_img_001.png" rel="lightbox-demo" title="my caption"><br /> | ||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/f/f8/2017_HUST_China_interlab_img_001.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> | ||
+ | </a> | ||
+ | <div class="col-xs-12" > | ||
+ | <h5><strong> Part 2: Prepare the serial dilutions of Fluorescein </strong> </h5> | ||
+ | <ul style="list-style: none; font-size:10px;"> | ||
+ | <li>①Add 100 μl of 1xPBS into wells A2, B2, C2, D2....A12, B12, C12, D12.</li> | ||
+ | <li>②Add 200 μl of 1x Fluorescein stock solution into A1, B1, C1, D1.</li> | ||
+ | <li>③ Transfer 100 μl of Fluorescein stock solution from A1 into A2.</li> | ||
+ | <li>④ Mix A2 by pipetting up and down 3x and transfer 100 μl into A3.</li> | ||
+ | <li>⑤ Mix A3 by pipetting up and down 3x and transfer 100 μl into A4.</li> | ||
+ | <li>⑥ Mix A4 by pipetting up and down 3x and transfer 100 μl into A5.</li> | ||
+ | <li>⑦ Mix A5 by pipetting up and down 3x and transfer 100 μl into A6.</li> | ||
+ | <li>⑧ Mix A6 by pipetting up and down 3x and transfer 100 μl into A7.</li> | ||
+ | <li>⑨ Mix A7 by pipetting up and down 3x and transfer 100 μl into A8.</li> | ||
+ | <li>⑩ Mix A8 by pipetting up and down 3x and transfer 100 μl into A9.</li> | ||
+ | <li>⑪ Mix A9 by pipetting up and down 3x and transfer 100 μl into A10.</li> | ||
+ | <li>⑫ Mix A10 by pipetting up and down 3x and transfer 100 μl into A11.</li> | ||
+ | <li>⑬ Mix A11 by pipetting up and down 3x and transfer 100 μl into liquid waste.</li> | ||
+ | <li>⑭ Take care not to continue serial dilution into column 12.</li> | ||
+ | <li>⑮ Repeat dilution series for rows B, C, D. | ||
+ | <li>⑯ Measure fluorescence of all samples in all standard measurement modes in instrument. | ||
</ul> | </ul> | ||
− | |||
+ | </div> | ||
+ | |||
+ | </div> | ||
+ | <div id="section6" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | ||
+ | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px;margin-top:0;">Cell Culture Setup and Measurement</h3> | ||
+ | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/> | ||
+ | <a href="https://static.igem.org/mediawiki/2017/e/e6/2017_HUST_China_interlab_img_002.png" rel="lightbox-demo" title="my caption"><br /> | ||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/e/e6/2017_HUST_China_interlab_img_002.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> | ||
+ | </a> | ||
+ | <div class="col-xs-12"> | ||
+ | <h5><strong>Transformation:</strong> </h5> | ||
+ | <ul class="yuandian"> | ||
+ | <li> control</li> | ||
+ | <li> Negative control</li> | ||
+ | <li>Test Device 1: J23101+I13504</li> | ||
+ | <li>Test Device 2: J23106+I13504</li> | ||
+ | <li>Test Device 3: J23117+I13504</li> | ||
+ | <li>Test Device 4: J23101.BCD2.E0040.B0015</li> | ||
+ | <li>Test Device 5: J23106.BCD2.E0040.B0015</li> | ||
+ | <li>Test Device 6: J23117.BCD2.E0040.B0015</li> | ||
+ | </ul> | ||
+ | <h5><strong>Cell growth:</strong> </h5> | ||
+ | <li class="dropdown" style="list-style-type:none;"> | ||
+ | <button href="#" class="dropdown-toggle" data-toggle="dropdown"> | ||
+ | More details <b class="caret"></b> | ||
+ | </button> | ||
+ | <ul class="dropdown-menu" style="padding: 10px; background:#ccc;"> | ||
+ | <li> ① Pick 2 colonies from each of plate and inoculate each in 5-10 mL LB medium + Chloramphenicol (For antibiotic concentrations, please follow these guidelines:<a href="http://parts.igem.org/Help:Protocols/Antibiotic_Stocks">http://parts.igem.org/Help:Protocols/Antibiotic_Stocks). | ||
+ | </a> | ||
+ | </li> | ||
+ | <li>② Grow the cells overnight (16-18 hours) at 37°C and 220 rpm.</li> | ||
+ | <li>③ With 8 total devices (including controls) and 2 colonies per device, you should have 16 cultures.</li> | ||
+ | |||
+ | </ul> | ||
+ | </li> | ||
+ | <h5><strong>Cell growth, sampling, and assay</strong> </h5> | ||
+ | <li class="dropdown" style="list-style-type:none;"> | ||
+ | <button href="#" class="dropdown-toggle" data-toggle="dropdown"> | ||
+ | More details <b class="caret"></b> | ||
+ | </button> | ||
+ | <ul class="dropdown-menu" style="padding: 10px; background:#ccc;"> | ||
+ | |||
+ | <li>① Set your instrument to read Abs600 (as OD calibration setting)</li> | ||
+ | <li>② Measure Abs600 of the overnight cultures</li> | ||
+ | <li>③ Dilute the cultures to a target Abs600 of 0.02 (see the volume of preloading culture and media in Excel Dilution Calculation sheets) in 12 ml LB medium + Chloramphenicol in 50 mL falcon tube.</li> | ||
+ | <li>④ Remove 500uL of each culture for your zero time point and hold these samples on ice. (You should have 16 sample tubes from the time=0 time point)</li> | ||
+ | <li>⑤ Incubate the cultures at 37°C and 220 rpm.</li> | ||
+ | <li>⑥ Remove 500uL samples of each culture after 2, 4 and 6 hours of growth. Keep samples on ice while completing the additional time points. You should have 16 sample tubes for each time point.</li> | ||
+ | <li>⑦ Set up your four measurement plates: For colony #1 culture, pipet 100 uL samples into wells A, B, C and D of the row for that device. For colony #2, pipet 100uL samples into wells E, F, G, and H of the row for that device.</li> | ||
+ | <li>⑧ Read your plates, taking care to use the exact same settings used for your fluorescein measurement.</li> | ||
+ | |||
+ | |||
+ | </ul> | ||
+ | </li> | ||
+ | <h5><strong>The initial OD600 measurement of our overnight cultures.</strong> </h5> | ||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/1/1a/2017_HUST_China_interlab_img_table1.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> | ||
+ | </div> | ||
+ | <div class="col-xs-12"> | ||
+ | <h5><strong>What type of media did you use for this step? </strong> </h5> | ||
+ | <p>Luria Bertani </p> | ||
+ | <h5><strong>What type of vessel or container did you use to grow your cells? </strong> </h5> | ||
+ | <p>50 ml Falcon tube</p> | ||
+ | <h5><strong>What temperature setting did you use during the measurement?</strong> </h5> | ||
+ | <p>22℃</p> | ||
+ | <h5><strong>What type of 96-well plates did you use?</strong> </h5> | ||
+ | <p></p> | ||
+ | <h5><strong>Black plates with transparent/clear bottom (preferred)</strong> </h5> | ||
+ | <p>Flat</p> | ||
+ | <h5><strong>Measurement </strong> </h5> | ||
+ | <ul class="yuandian"> | ||
+ | <li>Measure OD and fluorescence of all samples</li> | ||
+ | </ul> | ||
+ | <h5><strong>Suggested Plate Layout for 96-well Plate</strong> </h5> | ||
+ | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/> | ||
+ | <a href="https://static.igem.org/mediawiki/2017/8/87/2017_HUST_China_interlab_img_003.png" rel="lightbox-demo" title="my caption"><br /> | ||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/8/87/2017_HUST_China_interlab_img_003.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> | ||
+ | </a> | ||
+ | </div> | ||
</div> | </div> | ||
+ | <div id="section7" style="border: solid 1px #666; margin:5px; padding:5px;border-radius:10px;overflow: hidden;"> | ||
+ | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px;margin-top:0;">Interlab Resultn</h3> | ||
+ | <h5 class="col-xs-12"><strong>OD600 Reference Point</strong> </h5> | ||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/5/52/2017_HUST_China_interlab_img_table2.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> | ||
+ | <h5 class="col-xs-12"><strong>Fluorescence standard curve</strong> </h5> | ||
+ | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/> | ||
+ | <a href="https://static.igem.org/mediawiki/2017/a/a1/2017_HUST_China_interlab_img_table3.png" rel="lightbox-demo" title="my caption"><br /> | ||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/a/a1/2017_HUST_China_interlab_img_table3.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> | ||
+ | </a> | ||
+ | <p class="col-xs-12"><strong>NOTE: 50uM Sample exceeds the range of measurements</strong> </p> | ||
+ | <h5 class="col-xs-12"><strong>OD600 Reference Point</strong> </h5> | ||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/d/d1/2017_HUST_China_interlab_img_table4.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> | ||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/d/df/2017_HUST_China_interlab_img_table5.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> | ||
+ | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/> | ||
+ | <a href="https://static.igem.org/mediawiki/2017/b/b3/2017_HUST_China_interlab_img_table6.png" rel="lightbox-demo" title="my caption"><br /> | ||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/b/b3/2017_HUST_China_interlab_img_table6.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> | ||
+ | </a> | ||
+ | <p class="col-xs-12"><strong>Final scaling level determined from medium-high points likely to be less impacted by saturation or pipetting error.</strong> </p> | ||
+ | <p class="col-xs-12"><strong>If needed, you can shift which points are used, but it is likely better to correct instrument settings and protocol.</strong> </p> | ||
+ | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/> | ||
+ | <a href="https://static.igem.org/mediawiki/2017/b/b8/2017_HUST_China_interlab_img_table7.png" rel="lightbox-demo" title="my caption"><br /> | ||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/b/b8/2017_HUST_China_interlab_img_table7.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> | ||
+ | </a> | ||
+ | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/> | ||
+ | <a href="https://static.igem.org/mediawiki/2017/e/ea/2017_HUST_China_interlab_img_table8.png" title="my caption"><br /> | ||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/e/ea/2017_HUST_China_interlab_img_table8.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> | ||
+ | </a> | ||
+ | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/> | ||
+ | <a href="https://static.igem.org/mediawiki/2017/d/d2/2017_HUST_China_interlab_img_table9.png" rel="lightbox-demo" title="my caption"><br /> | ||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/d/d2/2017_HUST_China_interlab_img_table9.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> | ||
+ | </a> | ||
+ | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/> | ||
+ | <a href="https://static.igem.org/mediawiki/2017/2/2b/2017_HUST_China_interlab_img_table10.png" rel="lightbox-demo" title="my caption" style="padding: 10px;"><br /> | ||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/2/2b/2017_HUST_China_interlab_img_table10.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> | ||
+ | </a> | ||
+ | |||
+ | </div> | ||
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Latest revision as of 18:00, 1 November 2017