Team:Aachen/TestBench2

From 2014.igem.org

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__TOC__
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<table class="transbg" width="100%"  cellspacing="0" height="500px">
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<table class="transbg" width="975px"  cellspacing="0">
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= Media =
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<tr>
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== LB medium ==
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<td width="95%" align="center">
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# weight components
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<!--====================================================================-->
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::* '''5 g/L NaCl'''
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<!--====================================================================-->
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::* '''10 g/L tryptone'''
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<!--======================PUT CONTENT HERE==============================-->
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::* '''5 g/L yeast extract'''
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<!--======================vvvvvvvvvvvvvvvv==============================-->
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::* (15 g/L agar for plates)
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<!--======================vvvvvvvvvvvvvvvv==============================-->
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# fill up to 1&nbsp;L with deionized water
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<div class="shadow">
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# '''mix well''' by shaking
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<img src="http://2014.igem.org/wiki/images/0/05/Aachen_TeamfotoApril01.jpg" width="100%">
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# autoclave
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</div>
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## autoclaving tape, caps slightly unscrewed
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## base of the pot has to be covered with deionized water
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## close lid
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## heat '''level 3 until the pressure valve opens'''
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## reduce '''heat level to 1.5'''
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## set timer to '''20 minutes'''
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## turn heater off
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## '''wait until the pressure valve retracts''' (30-45 minutes)
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## open, close caps & shake
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# for plates, wait until you can touch the bottle ('''<60 °C''', clean bench!)
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# add antibiotics and '''shake''' (gloves!)
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<h3>Welcome!<br> We are iGEM Team Aachen 2014 </h3>
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== Hartmans minimal medium (HM) ==
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<p> If you want to find out more about Cellock Holmes, scroll down! </p>
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= Agar Chips =
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= Transformation =
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== Heat Shock ==
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# thaw cells on ice
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<table class="transbg" width="100%"  cellspacing="0" height="500px">
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# add 1&nbsp;µl of plasmid DNA
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<tr>
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# incubate on ice for 30'
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<td></td>
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# heat shock at 42&nbsp;°C for 60''
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<td width="975px" align="center">
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# incubate on ice for 5'
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# add 200&nbsp;µl of SOC media
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# incubate at 37&nbsp;°C for 2&nbsp;h
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# plate 20&nbsp; and 200&nbsp;µl on plates with the appropiate antibiotic
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<table class="transbg" width="975px"  cellspacing="0">
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== Electroporation ==
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<img src="http://2014.igem.org/wiki/images/0/0a/DwikitemplateB_dnastrand.png">
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===What is ''Cellock Holmes'' about?===
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Many young scientists dream about presenting their first big research project to a renowned professional audience. The prestigious iGEM competition provides us with an opportunity to make this dream come true. iGEM offers the ideal chance to expand our knowledge in different areas outside of our program, and to directly apply this knowledge to a current problem. Furthermore, we have the opportunity to learn modern research techniques, which go beyond the scope of our university studies. Working in an interdisciplinary team forces is to broaden our horizons, and we can learn from students studying different fields. Moreover, the competition will allow us to improve important soft skills such as the ability to work in a team, time and project management, and maintaining motivation throughout the project. We are looking forward to the international atmosphere of iGEM, and to meet like-minded student from all over the world. Lastly, it is the enthusiasm for fancy developments in synthetic biology that not only lead to new innovative products and a better life standard, but also to more sustainability.
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[[Team:Aachen]]
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= PCR =
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Within the scope of our project, we want to develop a system with which pathogens on solid surfaces can be detected quickly and cost-effectively. This is essential in many respects, because presently there are no effective analytic methods for use in the health sector that can meet the increasing demands of affordability and speed. It is particularly important to detect microorganisms on solid surfaces in places where good hygiene is crucial, since—even after cleaning—microorganisms can still be present in dangerous amounts. This is demonstrated by the high number of 1.7 million infections per year in the US health sector, of which approximately 10,000 resulted in death (Klevens et al., 2002). A large number of these cases would be preventable, however, there is a present lack of adequate tools for practical application. We not only want to develop a detection system that alerts the patient to the presence of pathogens, but one that can also identify and quantify these microorganisms. Only in this way is it possible to reliably estimate the danger of infection.
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== Colony PCR ==
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</html>{{Team:Aachen/JumpUp}}<html>
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</table>
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</td>
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== QuikChange ==
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<td></td>
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= Clonings =
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== Restriction Digest ==
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<table class="transbg" width="100%"  cellspacing="0">
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== Ligation ==
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Standard: <b>HTML-formatting</b></br>
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'''Wiki-formatting''' inside of these tags
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</html>{{Team:Aachen/JumpUp}}<html>
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== Gibson Assembly ==
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<td></td>
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<h3> Who will your project help?</h3>
 
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<p> Tell your audience how your project will help the environment, science, medicine or everything else.</p>
 
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<h3>Why did you choose this project?</h3>
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<p> What motivated you to work on this project? Tell us what inspires you.</p>
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Revision as of 21:29, 17 September 2014


Contents


Media

LB medium

  1. weight components
  • 5 g/L NaCl
  • 10 g/L tryptone
  • 5 g/L yeast extract
  • (15 g/L agar for plates)
  1. fill up to 1 L with deionized water
  2. mix well by shaking
  3. autoclave
    1. autoclaving tape, caps slightly unscrewed
    2. base of the pot has to be covered with deionized water
    3. close lid
    4. heat level 3 until the pressure valve opens
    5. reduce heat level to 1.5
    6. set timer to 20 minutes
    7. turn heater off
    8. wait until the pressure valve retracts (30-45 minutes)
    9. open, close caps & shake
  4. for plates, wait until you can touch the bottle (<60 °C, clean bench!)
  5. add antibiotics and shake (gloves!)

Hartmans minimal medium (HM)

Agar Chips

Transformation

Heat Shock

  1. thaw cells on ice
  2. add 1 µl of plasmid DNA
  3. incubate on ice for 30'
  4. heat shock at 42 °C for 60
  5. incubate on ice for 5'
  6. add 200 µl of SOC media
  7. incubate at 37 °C for 2 h
  8. plate 20  and 200 µl on plates with the appropiate antibiotic

Electroporation

PCR

Colony PCR

QuikChange

Clonings

Restriction Digest

Ligation

Gibson Assembly