Difference between revisions of "Team:NPU-China/InterLab"

 
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                 <button type="button" class="navbar-toggle" data-toggle="collapse" data-target="#bs-example-navbar-collapse-1"> <span class="sr-only">Toggle navigation</span> <span class="icon-bar"></span> <span class="icon-bar"></span> <span class="icon-bar"></span> </button>
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                 <button type="button" class="navbar-toggle" data-toggle="collapse" data-target="#bs-example-navbar-collapse-1">
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                    <span class="sr-only">Toggle navigation</span>
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                 <a class="navbar-brand" href="https://2017.igem.org/Team:NPU-China">
 
                 <a class="navbar-brand" href="https://2017.igem.org/Team:NPU-China">
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                  <img src="https://static.igem.org/mediawiki/2017/2/29/NPU-logo.png" style="max-width:50px;margin-top:-10px;">
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                    <img src="https://static.igem.org/mediawiki/2017/2/29/NPU-logo.png" style="max-width:50px;margin-top:-10px;">
 
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                 <ul class="nav navbar-nav navbar-right">
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                     <li class="active"> <a href="https://2017.igem.org/Team:NPU-China">Home</a> </li>
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                     <li>
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                     <li class="dropdown"> <a href="#" class="dropdown-toggle" data-toggle="dropdown">Team<b class="caret"></b></a>
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                        <a href="https://2017.igem.org/Team:NPU-China">Home</a>
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                    </li>
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                     <li class="dropdown">
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                        <a href="#" class="dropdown-toggle" data-toggle="dropdown">Team
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                            <b class="caret"></b>
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                         <ul class="dropdown-menu">
 
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                             <li> <a href="https://2017.igem.org/Team:NPU-China/Aboutus">About us</a> </li>
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                             <li>
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                             <li> <a href="https://2017.igem.org/Team:NPU-China/Attributions">Attributions</a> </li>
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                                <a href="https://2017.igem.org/Team:NPU-China/Aboutus">About us</a>
 +
                            </li>
 +
                             <li>
 +
                                <a href="https://2017.igem.org/Team:NPU-China/Attributions">Attributions</a>
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                            </li>
 
                         </ul>
 
                         </ul>
 
                     </li>
 
                     </li>
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                     <li class="dropdown"> <a href="#" class="dropdown-toggle" data-toggle="dropdown">Project<b class="caret"></b></a>
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                     <li class="dropdown">
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                        <a href="#" class="dropdown-toggle" data-toggle="dropdown">Project
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                            <b class="caret"></b>
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                        </a>
 
                         <ul class="dropdown-menu">
 
                         <ul class="dropdown-menu">
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                             <li> <a href="https://2017.igem.org/Team:NPU-China/Background">Background</a> </li>
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                             <li>
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                             <li> <a href="https://2017.igem.org/Team:NPU-China/Description">Description</a> </li>
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                                <a href="https://2017.igem.org/Team:NPU-China/Background">Background</a>
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                             <li> <a href="https://2017.igem.org/Team:NPU-China/Design">Design</a> </li>
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                            </li>
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                             <li> <a href="https://2017.igem.org/Team:NPU-China/Model">Model</a> </li>
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                             <li>
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                             <li> <a href="https://2017.igem.org/Team:NPU-China/Proofofconcept">Proof of concept</a> </li>
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                                <a href="https://2017.igem.org/Team:NPU-China/Description">Description</a>
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                             <li> <a href="https://2017.igem.org/Team:NPU-China/Demonstrate">Demonstrate</a> </li>
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                            </li>
 +
                             <li>
 +
                                <a href="https://2017.igem.org/Team:NPU-China/Design">Design</a>
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                            </li>
 +
                             <li>
 +
                                <a href="https://2017.igem.org/Team:NPU-China/Model">Model</a>
 +
                            </li>
 +
                             <li>
 +
                                <a href="https://2017.igem.org/Team:NPU-China/Proofofconcept">Proof of concept</a>
 +
                            </li>
 +
                             <li>
 +
                                <a href="https://2017.igem.org/Team:NPU-China/Demonstrate">Demonstrate</a>
 +
                            </li>
 
                         </ul>
 
                         </ul>
 
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                     </li>
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                     <li class="dropdown"> <a href="#" class="dropdown-toggle" data-toggle="dropdown">Parts<b class="caret"></b></a>
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                     <li class="dropdown">
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                        <a href="#" class="dropdown-toggle" data-toggle="dropdown">Parts
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                            <b class="caret"></b>
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                         <ul class="dropdown-menu">
 
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                             <li> <a href="https://2017.igem.org/Team:NPU-China/BasicParts">Basic Parts</a> </li>
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                             <li>
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                             <li> <a href="https://2017.igem.org/Team:NPU-China/CompositeParts">Composite Parts</a> </li>
+
                                <a href="https://2017.igem.org/Team:NPU-China/BasicParts">Basic Parts</a>
 +
                            </li>
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                             <li>
 +
                                <a href="https://2017.igem.org/Team:NPU-China/CompositeParts">Composite Parts</a>
 +
                            </li>
 
                         </ul>
 
                         </ul>
 
                     </li>
 
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                     <li> <a href="https://2017.igem.org/Team:NPU-China/Hardware">Hardware</a> </li>
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                     <li class="dropdown"> <a href="#" class="dropdown-toggle" data-toggle="dropdown">HP<b class="caret"></b></a>
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                        <a href="https://2017.igem.org/Team:NPU-China/Hardware">Hardware</a>
 +
                    </li>
 +
                     <li class="dropdown">
 +
                        <a href="#" class="dropdown-toggle" data-toggle="dropdown">HP
 +
                            <b class="caret"></b>
 +
                        </a>
 
                         <ul class="dropdown-menu">
 
                         <ul class="dropdown-menu">
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                             <li> <a href="https://2017.igem.org/Team:NPU-China/HP/Silver">Silver</a> </li>
+
                             <li>
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                             <li> <a href="https://2017.igem.org/Team:NPU-China/HP/Gold_Integrated">Gold</a> </li>
+
                                <a href="https://2017.igem.org/Team:NPU-China/HP/Silver">Silver</a>
 +
                            </li>
 +
                             <li>
 +
                                <a href="https://2017.igem.org/Team:NPU-China/HP/Gold_Integrated">Gold</a>
 +
                            </li>
 
                         </ul>
 
                         </ul>
 
                     </li>
 
                     </li>
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                     <li> <a href="https://2017.igem.org/Team:NPU-China/Collaborations">Collaborations</a> </li>
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                     <li>
−
                     <li> <a href="https://2017.igem.org/Team:NPU-China/Achievements">Achievements</a> </li>
+
                        <a href="https://2017.igem.org/Team:NPU-China/Collaborations">Collaborations</a>
−
                     <li> <a href="https://2017.igem.org/Team:NPU-China/InterLab">InterLab</a> </li>
+
                    </li>
 +
                     <li>
 +
                        <a href="https://2017.igem.org/Team:NPU-China/Achievements">Achievements</a>
 +
                    </li>
 +
                     <li class="active">
 +
                        <a href="https://2017.igem.org/Team:NPU-China/InterLab">InterLab</a>
 +
                    </li>
  
−
                     <li class="dropdown"> <a href="#" class="dropdown-toggle" data-toggle="dropdown">Notebook<b class="caret"></b></a>
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                     <li class="dropdown">
 +
                        <a href="#" class="dropdown-toggle" data-toggle="dropdown">Notebook
 +
                            <b class="caret"></b>
 +
                        </a>
 
                         <ul class="dropdown-menu">
 
                         <ul class="dropdown-menu">
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                             <li> <a href="https://2017.igem.org/Team:NPU-China/Labnotes">Labnotes</a> </li>
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                             <li>
−
                             <li> <a href="https://2017.igem.org/Team:NPU-China/Protocols">Protocols</a> </li>
+
                                <a href="https://2017.igem.org/Team:NPU-China/Labnotes">Labnotes</a>
 +
                            </li>
 +
                             <li>
 +
                                <a href="https://2017.igem.org/Team:NPU-China/Protocols">Protocols</a>
 +
                            </li>
 
                         </ul>
 
                         </ul>
 
                     </li>
 
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     <div class="batu" style="background: url('https://static.igem.org/mediawiki/2017/f/fe/Npu-background.png') no-repeat fixed; overflow: hidden;">
 
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         <!-- Page Content -->
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         <img class="img-responsive" src="https://static.igem.org/mediawiki/2017/a/a6/%E9%A2%98%E7%9B%AE%E5%B0%8F%E9%80%9A%E6%A0%8Finterlab.jpg">
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                     <h2 class="page-header" align="center" >Abstract</h2>
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                     <h2 style="text-align:center">Overview</h2>
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                     <h4>Acrylic acid is a bulk chemical raw material, which is widely used in many fields because of its excellent
+
                     <h4>The aim of the interlab this year was to explore--
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                         polymerization capacity, such as paint, glue, and even mobile phone screen protective film. The average
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                        <br> "How close can the numbers be when fluorescence is measured all around the world?"
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                         annual market demand of acrylic acid is up to 8 million tons, and the market value is nearly 10 billion
+
                        <br> Participating in the interlab was a valuable experience, and it allowed us to make use of technology
−
                         US dollars. It has broad market prospect. At present, acrylic acid is made from propylene (which
+
                         we had at our disposal that otherwise wasn’t relevant to our project. It added an extra dimension
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                         is obtained by petroleum cracking) after multi-step treatment. The production process causes pollution,
+
                         to our lab work, and was a lot of fun!
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                         high energy consumption and it is unsustainable.<br> This year, we aim to use a green and environmentally
+
                         <br>
−
                         friendly carbon source, glycerol to achieve all green production of acrylic acid. Compared to traditional
+
                        <br> 1. OD600 reference point
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                         chemical synthesis methods, Synbio is green and sustainable, and glycerol is cheaper than ethylene.
+
                         <div align="center">
 +
                            <img src="https://static.igem.org/mediawiki/2017/c/c0/Interlab-1.1.png" class="img-responsive" width="40%" height="40%">
 +
                        </div>
 +
                         <br>
 +
                         <br>
 +
                        <br> 2. Fluorescein standard curve
 +
                         <div class="col-md-12">
 +
                            <div class="col-md-6">
 +
                                <img src="https://static.igem.org/mediawiki/2017/9/91/Interlab-1.2.png" class="img-responsive">
 +
                            </div>
 +
                            <div class="col-md-6">
 +
                                <img src="https://static.igem.org/mediawiki/2017/a/a0/Interlab-2.1.png" class="img-responsive">
 +
                            </div>
 +
                        </div>
 
                     </h4>
 
                     </h4>
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                </div>
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                    <br>
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                    <br>
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            <!-- Marketing Icons Section -->
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                        <h4> 3. Raw Plate Reader Measurements</h4>
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                <div class="col-md-12">
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                    </div>
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                    <h2 class="page-header" align="center">We construct cell factory based on 4 levels, which are—</h2>
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                    <div class="col-md-12">
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                        <div class="col-md-6">
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                            <img src="https://static.igem.org/mediawiki/2017/6/6d/Interlab_%281%29.jpg" class="img-responsive">
 +
                        </div>
 +
                        <div class="col-md-6">
 +
                            <img src="https://static.igem.org/mediawiki/2017/d/dd/Interlab_%282%29.jpg" class="img-responsive">
 +
                        </div>
 +
                    </div>
 +
                    <br>
 
                     <br>
 
                     <br>
−
                </div>
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            </div>
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                        <div class="col-md-6">
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                            <img src="https://static.igem.org/mediawiki/2017/9/91/Interlab_%283%29.jpg" class="img-responsive">
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                            <img src="https://static.igem.org/mediawiki/2017/f/f5/Interlab_%284%29.jpg" class="img-responsive">
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 +
                    <br>
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                    <div style="padding-top:600px"></div>
 +
                        <h2 style="text-align:center" >Observation</h2>
 +
                        <h4>
 +
                            1. It can be easily observed that Device1 and Device 4 didn’t grow after 6 hours. In fact, Device1 express high level of
 +
                            GFP after being cultured all night long.
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                            <br>
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                            <br> 2.In Fluorescein standard curve, it’s not linear when the concentration is high. But the Fluorescein
 +
                            in Raw Plate Reader Measurements doesn’t reach the the non-linear region so the standard curve
 +
                            is believable.
 +
                            <br>
 +
                            <br> 3. It was found there was great variation in readings when using different settings on the plate
 +
                            reader. Given that iGEM HQ has kept the protocol consistent, it is suggested that the desired
 +
                            plate reader settings are also specified to ensure consistency throughout the results.
 +
                            <br>
 +
                            <br> 4. The dilution forum provided by iGEM foundation is useful. After dilution, almost every OD600
 +
                            is around 0.04.
 +
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                    <h4>We use ceaS2 enzyme as the core part, but acrylic acid is a byproduct of ceaS2 enzyme, the wild type
 
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                        's catalytic effect is very weak, whose production is only 1mg/L. So we hope to improve the catalytic
 
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                        effect of ceaS2 enzyme.<br> We designed ceaS2 enzyme mutants via the AEMD(Auto Enzyme Mutation Design)
 
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                        platform and screened for better-worked ceaS2 mutants by HPLC(High Performance Liquid Chromatography)
 
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                        and HTS(High throughput screening).
 
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                    <h4>Respectively, E. coli and S. cerevisiae are the two sorts of model organisms that are most convenient
 
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                        to operate in the prokaryote and eukaryote. Therefore, in terms of our choice of the chassis organisms,
 
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                        we have them both tested, which were E. coli MG1655 and S. cerevisiae BY4741 individually.
 
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                    <h4>We need to design two different metabolic pathways for two different chassis organisms and propose different
 
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                        optimization schemes for them.We introduced the ceaS2 enzyme exogenously on the basis of the glycerol
 
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                        metabolism of the two bacteria, so that it could produce the target product acrylic acid using the
 
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                        intermediates G3P and DHAP.Besides having finished the construction of the pathways, we also reconstructed
 
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                        and optimized the original metabolic pathway to increase the carbon flux rate of the designed pathway
 
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                        and reduce the loss of bypass carbon flux.</h4>
 
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                    <h4>All of the previous processes are to build the engineering strains which have a high production of acrylic
 
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                        acid that we need. In the subsequent fermentation, we also need to determine the best parameters
 
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                        of the engineering strain.<br> Therefore, we selected the carbon source, Buffer, temperature, pH
 
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                        and other conditions to optimize the cell production process.</h4>
 
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Latest revision as of 03:19, 31 October 2017

Overview

The aim of the interlab this year was to explore--
"How close can the numbers be when fluorescence is measured all around the world?"
Participating in the interlab was a valuable experience, and it allowed us to make use of technology we had at our disposal that otherwise wasn’t relevant to our project. It added an extra dimension to our lab work, and was a lot of fun!

1. OD600 reference point



2. Fluorescein standard curve



3. Raw Plate Reader Measurements







Observation

1. It can be easily observed that Device1 and Device 4 didn’t grow after 6 hours. In fact, Device1 express high level of GFP after being cultured all night long.


2.In Fluorescein standard curve, it’s not linear when the concentration is high. But the Fluorescein in Raw Plate Reader Measurements doesn’t reach the the non-linear region so the standard curve is believable.

3. It was found there was great variation in readings when using different settings on the plate reader. Given that iGEM HQ has kept the protocol consistent, it is suggested that the desired plate reader settings are also specified to ensure consistency throughout the results.

4. The dilution forum provided by iGEM foundation is useful. After dilution, almost every OD600 is around 0.04.