Difference between revisions of "Team:Kent/Attributions"

 
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                     <ul class="drop-menu menu-1">
 
                     <ul class="drop-menu menu-1">
 
                         <a href="https://2017.igem.org/Team:Kent/Description"><li>Description</li></a>
 
                         <a href="https://2017.igem.org/Team:Kent/Description"><li>Description</li></a>
−
<a href="https://2017.igem.org/Team:Kent/Design"><li> Design </li></a>
+
<a href="https://2017.igem.org/Team:Kent/Model"><li>Modelling</li></a>
 
                       <a href="https://2017.igem.org/Team:Kent/Results"><li>Results</li></a>
 
                       <a href="https://2017.igem.org/Team:Kent/Results"><li>Results</li></a>
−
                        <a href="https://2017.igem.org/Team:Kent/Model"><li>Modelling</li></a>
+
                     
−
<a href="https://2017.igem.org/Team:Kent/Demonstrate"><li>Demonstrate</li></a>
+
 
                     </ul>
 
                     </ul>
 
                 <li>
 
                 <li>
 
                     <a href="#">Parts</a>
 
                     <a href="#">Parts</a>
 
                     <ul class="drop-menu menu-2">
 
                     <ul class="drop-menu menu-2">
−
<a href="https://2017.igem.org/Team:Kent/Parts"> <li> Parts </li></a>
+
 
 
                         <a href="https://2017.igem.org/Team:Kent/Basic_Part"><li>Basic Parts</li></a>
 
                         <a href="https://2017.igem.org/Team:Kent/Basic_Part"><li>Basic Parts</li></a>
−
                        <a href="https://2017.igem.org/Team:Kent/Composite_Part"><li>Composite Parts</li></a>
+
                     
−
<a href = "https://2017.igem.org/Team:Kent/Part_Collection"><li> Part Collection </li></a>
+
  
 
                     </ul>
 
                     </ul>
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                     <ul class="drop-menu menu-2">
 
                     <ul class="drop-menu menu-2">
 
                         <a href="https://2017.igem.org/Team:Kent/Safety"><li>Project Safety</li></a>
 
                         <a href="https://2017.igem.org/Team:Kent/Safety"><li>Project Safety</li></a>
−
                         <a href="https://2017.igem.org/Team:Kent/Signs"><li>Hazard Signs</li></a>
+
                          
 
                     </ul>
 
                     </ul>
 
                 </li>
 
                 </li>
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                     <a href="#">Human Practices</a>
 
                     <a href="#">Human Practices</a>
 
                     <ul class="drop-menu menu-2">
 
                     <ul class="drop-menu menu-2">
−
                         <a href="https://2017.igem.org/Team:Kent/HP/Silver"><li>Silver</li></a>
+
                         <a href="https://2017.igem.org/Team:Kent/HP/Silver"><li>Integrate HP</li></a>
−
<a href="https://2017.igem.org/Team:Kent/HP/Gold_Integrated"><li>Gold</li></a>
+
 
                         <a href="https://2017.igem.org/Team:Kent/Engagement"><li>Public Engagement</li></a>
 
                         <a href="https://2017.igem.org/Team:Kent/Engagement"><li>Public Engagement</li></a>
 
                         <a href="https://2017.igem.org/Team:Kent/InterLab"><li>Interlab</li></a>
 
                         <a href="https://2017.igem.org/Team:Kent/InterLab"><li>Interlab</li></a>
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<img src = "https://static.igem.org/mediawiki/2017/thumb/9/9c/T--Kent--AttributionHeader2.png/637px-T--Kent--AttributionHeader2.png" id="header2">
 
<img src = "https://static.igem.org/mediawiki/2017/thumb/9/9c/T--Kent--AttributionHeader2.png/637px-T--Kent--AttributionHeader2.png" id="header2">
 
         </div>
 
         </div>
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</div>
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<header class="hull">
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−
<label for="acc-close" class="hull-title">Basic Protocols</label>
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<div id="textbox1" ><br><br>
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</header>
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<ul><li><Technical services led by Julian Cook- School of Biosciences, University of Kent<br>
−
<input type="radio" name="droptext" id="cb1" />
+
Technical services provided us with equipment and materials for use during our time in the lab for the duration of the project, and autoclaved our biological waste. They provided everything from petri dishes to ingredients to make things like gels and agar, and glassware that was essential to our lab work. Julian Cook, head of technical services, also gave us a safety induction before our lab work began.<br></li><br>
−
<section class="hull">
+
<li>Taylor Monaghan<br>
−
<label class="hull-title" for="cb1">Production of Lysogeny broth (LB)</label>
+
Taylor is a PhD student at the University of Kent who aided us in learning basic molecular biology techniques, acting as our lab supervisor. He guided us on safety in the lab,= and used his experience in the lab to provide a vital assistance to our team.<br></li><br>
−
<label class="hull-close" for="acc-close"></label>
+
<li>Dr Rosalyn Masterton<br>
−
<div class="hull-content">For 1 litre of LB a mixture of 10g of sodium chloride, 10g peptone, 5g of yeast extract as well as 1
+
    Dr Masterton kindly helped us with gaining understanding in molecular biology and cloning as well as offering us a sample of pcDNA 3.1 (+) mammalian vector for cloning our construct for imaging.<br></li><br>
−
litre of distilled water in a glass bottle. We then used a magnetic spinner to help mix the powders
+
<li>Dr Emma Mead<br>
−
with the water, we avoided shaking the glass bottle as it would cause froth and waste some of the
+
Dr Emma Mead assisted us with all the mammalian cell culturing procedures. She kindly offered us a sample of HEK293 cells and aided us in experiment design, passaging and transfection. She also advised us on future design tweaks and experiments    <br></li><br>
−
LB.
+
<li>Dr Alexandra Moores<br>
−
<br>
+
Dr Alexandra Moores was our point of contact in Dr Neil Kad’s lab for most procedures. She also advised us in the optimization of experiments and protocols to achieve better results<br></li><br>
−
When making the LB we also made another litre batch and added 15g of agar extract to be able to
+
    <li>
−
grow bacteria on plates.</div>
+
Professor Mark Smales<br>
−
</section>
+
    Professor Mark Smales offered lab space for mammalian cell culturing and the chemicals necessary for these experiments.<br></li><br>
−
<input type="radio" name="droptext" id="cb2" />
+
<li>Dr Neil Kad<br>
−
<section class="hull">
+
    Principal investigator and allowed us to work in his lab after we had to vacant the teaching lab space at the start of term.<br></li><br>
−
<label class="hull-title" for="cb2">Production of SOB medium and magnesium stock</label>
+
<li>Dr Peter Ellis<br>
−
<label class="hull-close" for="acc-close"></label>
+
    Dr Peter Ellis was an advisor who helped us perfect our project design with his knowledge on genetics<br></li><br>
−
<div class="hull-content">Bringing together 20g of tryptone, 5g of yeast extract, 0.584g of NaCl, 0.186g of KCl and mixing it
+
<li>Dr Wei-Feng Xue<br>
−
with 990ml of millipure water (using the magnetic mixer again) which was then put in to autoclave
+
    Dr Wei-Feng Xue used to be PI for iGEM teams of Kent in the past and advised us on the important points of iGEM and the requirements for the competition. <br></li><br>
−
to sterilise it, after it was taken out and let for it to cool down to below 60 o C.
+
<li>Dr Gary Robinson<br>
−
<br>
+
    Dr Gary Robinson was an advisor who helped us looking at the business side of the project and helped us form the future aspects of our project. He also advised on the possibilities with the Addgene purchases.<br></li><br>
−
10ml of 2M Mg 2+ stock was then added and then brought to 100ml with millipure water, 0.2mm
+
<li>Dr Richard Williamson<br>
−
filter sterilize was then used</div>
+
Dr Richard Williamson offered us a space to exhibit our project and ideas on the Open Day of the School of Biosciences where we could liaise with prospective undergraduate students. </li><br>
−
</section>
+
<li>
−
<input type="radio" name="droptext" id="cb3" />
+
Ian Brown<br>
−
<section class="hull">
+
Ian helped us with setting up the microscope and taking photos of our cells. He also gave us insight of how we might improve our design to enhance picture quality under the microscope.<br></li><br>
−
<label class="hull-title" for="cb3">Production of SOC medium and glucose stock</label>
+
 
−
<label class="hull-close" for="acc-close"></label>
+
<li>Snapgene<br>
−
<div class="hull-content">Once again bring 20g of tryptone, 5g of yeast of extract, 0.584g of NaCl, 0.186g of KCL, and then
+
Snapgene provided us with sponsorship for a free license for their plasmid visualisation software, which was crucial to the design of our biobrick and visualising any restriction sites, helping us carry out both restriction digests, ligations, and Gibson Assembly reactions, proving fundamental to the running of our project.<br></li><br>
−
bring 970 ml with millipure water and use the magnetic mixer once again, this was also then put in
+
 
−
to autoclave.
+
<li>Integrated DNA Technologies<br>
−
<br>
+
For providing us with 20kB of free DNA synthesis which we thoroughly took use of.<br></li><br>
−
10ml of 2M Mg 2+ stock and then bring it to 100ml with milllipure water, filter sterilize it with 0.2m
+
 
−
and then final add 20ml of 1M glucose stock.</div>
+
<li>School of Biosciences at the University of Kent<br>
−
</section>
+
For allowing us to work in teaching lab 1 over the summer and providing us with the equipment and expertise to enable us to compete in the igem competition in the first place. <br>
−
<input type="radio" name="droptext" id="acc-close" />
+
</li><br>
−
<input type="radio" name="droptext" id="cb4" />
+
 
−
<section class="hull">
+
</div>
−
<label class="hull-title" for="cb4">Production of Glycerol stock</label>
+
 
−
<label class="hull-close" for="acc-close"></label>
+
</div>
−
<div class="hull-content">If you wish to store bacteria long term, you will need to create a Glycerol Stock after
+
<div id="alldata">
−
inoculating an overnight liquid culture
+
<p></p>
−
<br>
+
</div>
−
<ul><li>Once bacterial growth has been achieved, 500μL of the overnight liquid
+
−
culture needs to be added to 500μL of 50% glycerol in a 2mL tube where it
+
−
should be gently mixed</li>
+
−
<li>The glycerol stock should then be frozen at -80 o C<ul>
+
−
<li> Successive freeze and thaw cycles will reduce the stocks shelf life</li></ul>
+
−
</li></ul></div>
+
−
</section>
+
−
<input type="radio" name="droptext" id="acc-close" />
+
−
<input type="radio" name="droptext" id="cb5" />
+
−
<section class="hull">
+
−
<label class="hull-title" for="cb5">Running Agarose Gel</label>
+
−
<label class="hull-close" for="acc-close"></label>
+
−
<div class="hull-content">After the cells have been miniprepped and the plasmid put through a restriction digest, the agarose gel can be run.
+
−
<br>
+
−
<ul><li>Make up some agarose. This is done by taking 0.5g of agarose powder and putting it in a
+
−
250ml sterile conical flask, with 50ml of TAE buffer, then microwaving it in small pulses (20
+
−
seconds then swirling it around) until it is dissolved. Don’t overheat it or it will evaporate too
+
−
much. Make up the evaporated volume to 50ml with distilled water.</li>
+
−
<li>Add 1 vial of cybersafe (ask technical services for a tube of it and add all of it)</li>
+
−
<li>Line the white sides of the tank with the agarose solution, to seal it and prevent leakage. Use
+
−
a p1000 pipette set to 1ml. Let it dry (about 5 mins max)</li>
+
−
<li>Then pour all the agarose/sybrsafe solution into the tank and put in the comb. Let it set and
+
−
solidify (maximum 30 mins)</li>
+
−
<li>When the gel has set, remove the comb from the tank (gently!) and then cover the whole
+
−
tank with TAE buffer, so there’s at least half a centimetre of TAE covering the gel.</li>
+
−
<li>Now, the samples need to be loaded. Load some DNA markers (ask technical services for a
+
−
tube of this and load the whole tube) into well 1( left hand side) and then choose what you
+
−
load into wells 2, 3, and 4 etc. (make sure you note what’s in each lane!)</li>
+
−
<li>Load all of your digests into the wells 2,3, and 4.</li>
+
−
<li>Plug into a power supply and put the cover on. Run for 40 mins to an hour at 80v. The amps
+
−
don’t matter.</li>
+
−
<li>Once the visible markers have reached the half way point of the tank, turn off the power
+
−
supply and drain the TAE buffer form the tank. Remove the gel with a spatula and place in a
+
−
UV imaging box. Take an image of the gel under UV light, save it onto a USB stick.</li></ul></div>
+
−
</section>
+
−
<input type="radio" name="droptext" id="acc-close" />
+
−
</nav>
+
  
 
         <div id="foot">
 
         <div id="foot">

Latest revision as of 03:23, 16 December 2017


Attribution


  • Technical services provided us with equipment and materials for use during our time in the lab for the duration of the project, and autoclaved our biological waste. They provided everything from petri dishes to ingredients to make things like gels and agar, and glassware that was essential to our lab work. Julian Cook, head of technical services, also gave us a safety induction before our lab work began.

  • Taylor Monaghan
    Taylor is a PhD student at the University of Kent who aided us in learning basic molecular biology techniques, acting as our lab supervisor. He guided us on safety in the lab,= and used his experience in the lab to provide a vital assistance to our team.

  • Dr Rosalyn Masterton
    Dr Masterton kindly helped us with gaining understanding in molecular biology and cloning as well as offering us a sample of pcDNA 3.1 (+) mammalian vector for cloning our construct for imaging.

  • Dr Emma Mead
    Dr Emma Mead assisted us with all the mammalian cell culturing procedures. She kindly offered us a sample of HEK293 cells and aided us in experiment design, passaging and transfection. She also advised us on future design tweaks and experiments

  • Dr Alexandra Moores
    Dr Alexandra Moores was our point of contact in Dr Neil Kad’s lab for most procedures. She also advised us in the optimization of experiments and protocols to achieve better results

  • Professor Mark Smales
    Professor Mark Smales offered lab space for mammalian cell culturing and the chemicals necessary for these experiments.

  • Dr Neil Kad
    Principal investigator and allowed us to work in his lab after we had to vacant the teaching lab space at the start of term.

  • Dr Peter Ellis
    Dr Peter Ellis was an advisor who helped us perfect our project design with his knowledge on genetics

  • Dr Wei-Feng Xue
    Dr Wei-Feng Xue used to be PI for iGEM teams of Kent in the past and advised us on the important points of iGEM and the requirements for the competition.

  • Dr Gary Robinson
    Dr Gary Robinson was an advisor who helped us looking at the business side of the project and helped us form the future aspects of our project. He also advised on the possibilities with the Addgene purchases.

  • Dr Richard Williamson
    Dr Richard Williamson offered us a space to exhibit our project and ideas on the Open Day of the School of Biosciences where we could liaise with prospective undergraduate students.

  • Ian Brown
    Ian helped us with setting up the microscope and taking photos of our cells. He also gave us insight of how we might improve our design to enhance picture quality under the microscope.

  • Snapgene
    Snapgene provided us with sponsorship for a free license for their plasmid visualisation software, which was crucial to the design of our biobrick and visualising any restriction sites, helping us carry out both restriction digests, ligations, and Gibson Assembly reactions, proving fundamental to the running of our project.

  • Integrated DNA Technologies
    For providing us with 20kB of free DNA synthesis which we thoroughly took use of.

  • School of Biosciences at the University of Kent
    For allowing us to work in teaching lab 1 over the summer and providing us with the equipment and expertise to enable us to compete in the igem competition in the first place.