Difference between revisions of "Team:Edinburgh/InterLab"

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                     <h2 class="section-heading">OVERVIEW</h2>
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                     <h2 class="section-heading">Protocol Section</h2>                  
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                    Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aenean posuere mattis risus ut sollicitudin. Mauris tempus et neque in maximus. Maecenas in velit nec lacus molestie ultrices. Morbi lacinia pellentesque porttitor. Nam nulla enim, congue vel nisl vel, lobortis tempus turpis. Sed semper quis odio et eleifend. Fusce eleifend tortor nec facilisis finibus. Sed mollis erat et sapien convallis, posuere cursus lectus iaculis. Pellentesque at augue varius, molestie nulla vel, fringilla nisl. Aenean id rhoncus ante. Pellentesque at urna sed tortor porttitor scelerisque. Mauris lacinia ligula maximus, pulvinar dolor vel, varius nisl. Suspendisse pellentesque augue ac ornare tempor. Praesent sit amet purus at dolor aliquam molestie nec vitae ipsumLorem ipsum dolor sit amet, consectetur adipiscing elit. Aenean posuere mattis risus ut sollicitudin. Mauris tempus et neque in maximus. Maecenas in velit nec lacus molestie ultrices. Morbi lacinia pellentesque porttitor. Nam nulla enim, congue vel nisl vel, lobortis tempus turpis. Sed semper quis odio et eleifend. Fusce eleifend tortor nec facilisis finibus. Sed mollis erat et sapien convallis, posuere cursus lectus iaculis. Pellentesque at augue varius, molestie nulla vel, fringilla nisl. Aenean id rhoncus ante. Pellentesque at urna sed tortor porttitor scelerisque. Mauris lacinia ligula maximus, pulvinar dolor vel, varius nisl. Suspendisse pellentesque augue ac ornare tempor. Praesent sit amet purus at dolor aliquam molestie nec vitae ipsum
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              SECTION I - LOREM IPSUM
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                    <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne">What protocol was followed for growing cells for measurement?</a>
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                    <p>The protocol supplied in the iGEM 2015 InterLab Protocol  form was followed with a few minor changes.
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                    <br>
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                    Each device and the two controls were streaked out onto LB agar plates supplemented with chloramphenicol and incubated at 37°C for 22 hours.  3 colonies from each plate were inoculated into 50ml Falcon tubes containing 10ml of LB media and chloramphenicol.  The tubes were incubated upright on a platform shaker in a room kept at 37°C for 16 hours.</p>
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                <h5>Lorem ipsum dolor sit amet?</h5>
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                    <a data-toggle="collapse" data-parent="#accordion" href="#collapseTwo">What controls were used?</a>
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                    <p>The positive control used was the recommended BBa_I20270, a chloramphenicol resistant GFP expression device in pSB1C3.
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<br>
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The negative control used consisted of cells transformed with BioBrick number BBa_R0040. This part consists of the promoter pTetR and should therefore have very low fluorescence</p>
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                    <a data-toggle="collapse" data-parent="#accordion" href="#collapseThree">What agar was used to grow the cells?</a>
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                    <p>The cells were grown on LB agar supplemented with 1000x chloramphenicol.</p>
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                    <a data-toggle="collapse" data-parent="#accordion" href="#collapseFour">How were the cells prepared for measurement?</a>
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                    <p>A colony of cells was inoculated into 10ml of liquid LB with 10µl of 1000x chloramphenicol. The cultures were grown in a 50ml falcon tube, upright on shaking platform at 275rpm in 37°C room overnight.
 
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For each device three colonies were inoculated to act as the three biological replicates.</p>
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              SECTION II - LOREM IPSUM
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             </a>
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                <h5>Lorem ipsum dolor sit amet?</h5>
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                    <a data-toggle="collapse" data-parent="#accordion" href="#collapseFive">How was the fluorescence of each device measured? (followed protocol)</a>
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                    <p>After the overnight incubation period of 16 hours the liquid cultures were removed from the 37°C room. 200µl from each culture was added to a 96-well plate. The initial optical density of each sample was measured at 600nm using a plate reader. All samples were then diluted appropriately so as to have an OD600 value within 5% of 0.5.  Two additional technical replicates for each of the biological replicates were set up at this point. The OD600 was measured again to ensure all samples had an optical density within 5% of 0.5 before the fluorescence measurements were taken.
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<br>
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To measure fluorescence, the cells were excited at a wavelength of 485nm and emitted light at 538nm as measured.
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<br>
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The values obtained for fluorescence were in relative fluorescence units.</p>
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                    <a data-toggle="collapse" data-parent="#accordion" href="#collapseSix">What instrument was used to measure the fluorescence of each device?</a>
  
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                    <p>A SpectaMax M5 plate reader was used to obtain the values of fluorescence.</p>
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<section id="about">
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    <div class="container">
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            <div class="row">
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                <div class="col-lg-12 text-center">
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                    <h2 class="section-heading">mRFP</h2>                   
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                </div>
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  </div>
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</section>
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<div class="container">
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                    <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne">1. What is HTML?</a>
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                    <p>HTML stands for HyperText Markup Language. HTML is the main markup language for describing the structure of Web pages. <a href="http://www.tutorialrepublic.com/html-tutorial/" target="_blank">Learn more.</a></p>
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                <h4 class="panel-title">
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                    <a data-toggle="collapse" data-parent="#accordion" href="#collapseTwo">2. What is Bootstrap?</a>
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                    <p>Bootstrap is a powerful front-end framework for faster and easier web development. It is a collection of CSS and HTML conventions. <a href="http://www.tutorialrepublic.com/twitter-bootstrap-tutorial/" target="_blank">Learn more.</a></p>
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                <h4 class="panel-title">
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                    <a data-toggle="collapse" data-parent="#accordion" href="#collapseThree">3. What is CSS?</a>
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            </div>
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            <div id="collapseThree" class="panel-collapse collapse">
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                <div class="panel-body">
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                    <p>CSS stands for Cascading Style Sheet. CSS allows you to specify various style properties for a given HTML element such as colors, backgrounds, fonts etc. <a href="http://www.tutorialrepublic.com/css-tutorial/" target="_blank">Learn more.</a></p>
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Revision as of 11:15, 8 September 2015

InterLab Study

Protocol Section

The protocol supplied in the iGEM 2015 InterLab Protocol form was followed with a few minor changes.
Each device and the two controls were streaked out onto LB agar plates supplemented with chloramphenicol and incubated at 37°C for 22 hours. 3 colonies from each plate were inoculated into 50ml Falcon tubes containing 10ml of LB media and chloramphenicol. The tubes were incubated upright on a platform shaker in a room kept at 37°C for 16 hours.

The positive control used was the recommended BBa_I20270, a chloramphenicol resistant GFP expression device in pSB1C3.
The negative control used consisted of cells transformed with BioBrick number BBa_R0040. This part consists of the promoter pTetR and should therefore have very low fluorescence

The cells were grown on LB agar supplemented with 1000x chloramphenicol.

A colony of cells was inoculated into 10ml of liquid LB with 10µl of 1000x chloramphenicol. The cultures were grown in a 50ml falcon tube, upright on shaking platform at 275rpm in 37°C room overnight.
For each device three colonies were inoculated to act as the three biological replicates.

After the overnight incubation period of 16 hours the liquid cultures were removed from the 37°C room. 200µl from each culture was added to a 96-well plate. The initial optical density of each sample was measured at 600nm using a plate reader. All samples were then diluted appropriately so as to have an OD600 value within 5% of 0.5. Two additional technical replicates for each of the biological replicates were set up at this point. The OD600 was measured again to ensure all samples had an optical density within 5% of 0.5 before the fluorescence measurements were taken.
To measure fluorescence, the cells were excited at a wavelength of 485nm and emitted light at 538nm as measured.
The values obtained for fluorescence were in relative fluorescence units.

A SpectaMax M5 plate reader was used to obtain the values of fluorescence.

mRFP

HTML stands for HyperText Markup Language. HTML is the main markup language for describing the structure of Web pages. Learn more.

Bootstrap is a powerful front-end framework for faster and easier web development. It is a collection of CSS and HTML conventions. Learn more.

CSS stands for Cascading Style Sheet. CSS allows you to specify various style properties for a given HTML element such as colors, backgrounds, fonts etc. Learn more.