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Team:LMU-Munich/Notebook/Protocols
From 2014.igem.org
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= Protocols = | = Protocols = | ||
| - | Our | + | Our Organism of Choice is ''Bacillus subtilis''. Listed below are the protocols we used in our lab. |
| - | If you have not worked with this | + | If you have not worked with this organism yet, we highly recommend visiting the award winning [https://2012.igem.org/Team:LMU-Munich/Lab_Notebook/Protocols wiki of the iGEM Team LMU-Munich 2012]. |
== Cloning == | == Cloning == | ||
*[[Media:LMU_Munich14_PCR.pdf|PCR Methods]] | *[[Media:LMU_Munich14_PCR.pdf|PCR Methods]] | ||
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*[[Media:LMU_Munich14_Competent_Cells.pdf|''E. coli'' Competent Cells ]] | *[[Media:LMU_Munich14_Competent_Cells.pdf|''E. coli'' Competent Cells ]] | ||
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*[[Media:LMU-Munich14 Cloning.pdf|Cloning]] | *[[Media:LMU-Munich14 Cloning.pdf|Cloning]] | ||
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*[[Media:LMU Munich14 Plasmid Extraction using Alkaline Lysis Method.pdf|Plasmid Extraction using Alkaline Lysis Method]] | *[[Media:LMU Munich14 Plasmid Extraction using Alkaline Lysis Method.pdf|Plasmid Extraction using Alkaline Lysis Method]] | ||
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*[[Media:LMU-Munich14 QuikChange Site Directed Mutagenesis.pdf|QuickChange Site Directed Mutagenesis]] | *[[Media:LMU-Munich14 QuikChange Site Directed Mutagenesis.pdf|QuickChange Site Directed Mutagenesis]] | ||
| - | + | *[[Media:LMU Munich14 FusionPCR.pdf|Fusion PCR]] | |
*[[Media:LMU Munich14 Golden Gate Cloning.pdf|Golden Gate Cloning]] | *[[Media:LMU Munich14 Golden Gate Cloning.pdf|Golden Gate Cloning]] | ||
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== Working with ''B. subtilis'' == | == Working with ''B. subtilis'' == | ||
*[[Media:LMU Munich14 Media.pdf|Media]] | *[[Media:LMU Munich14 Media.pdf|Media]] | ||
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*[[Media:LMU Munich14 Transformation of Bacillus subtilis.pdf|Transformation of ''Bacillus subtilis'']] | *[[Media:LMU Munich14 Transformation of Bacillus subtilis.pdf|Transformation of ''Bacillus subtilis'']] | ||
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*[[Media:LMU_Munich14_isolation_of_genomic_DNA_from_Bacillus_for_transformation.pdf|SC-Lyse for transformation]] | *[[Media:LMU_Munich14_isolation_of_genomic_DNA_from_Bacillus_for_transformation.pdf|SC-Lyse for transformation]] | ||
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== Working with ''S. pneumoniae'' == | == Working with ''S. pneumoniae'' == | ||
| - | *[[Media:LMU Munich14 Blood Agar (D-Agar).pdf| | + | *[[Media:LMU Munich14 Blood Agar (D-Agar).pdf|Blood Agar for ''S. pneumoniae'']] |
| + | *[[Media:LMU Munich14 Todd-Hewitt-Broth (THB).pdf|Todd-Hewitt-Broth for ''S. pneumoniae'']] | ||
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| + | == Protein Methods == | ||
| + | *[[Media:LMU Munich14 Overexpression and analysis of tagged NOX and peptides.pdf|Overexpression and analysis of tagged NOX and peptides]] | ||
| + | *[[Media:LMU Munich14 Adhesion of peptides to S.pneumoniae.pdf|Adhesion of peptides to ''S.pneumoniae'']] | ||
== Assays == | == Assays == | ||
*[[Media:LMU-Munich14 Luminescence Assay.pdf|Luminescence Plate Reader Assay]] | *[[Media:LMU-Munich14 Luminescence Assay.pdf|Luminescence Plate Reader Assay]] | ||
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*[[Media:LMU Munich14 FACS.pdf|Flow Cytometry Analysis]] | *[[Media:LMU Munich14 FACS.pdf|Flow Cytometry Analysis]] | ||
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*[[Media:LMU Munich14 Spot-on-lawn Assay.pdf|Spot-on-lawn Assay]] | *[[Media:LMU Munich14 Spot-on-lawn Assay.pdf|Spot-on-lawn Assay]] | ||
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{{Template:Team:LMU-Munich/Playground/footer}} | {{Template:Team:LMU-Munich/Playground/footer}} | ||
<html><script>initiateNavigation("labbook");</script></html> | <html><script>initiateNavigation("labbook");</script></html> | ||
Latest revision as of 22:02, 4 December 2014
Protocols
Our Organism of Choice is Bacillus subtilis. Listed below are the protocols we used in our lab. If you have not worked with this organism yet, we highly recommend visiting the award winning wiki of the iGEM Team LMU-Munich 2012.
Cloning
- PCR Methods
- E. coli Competent Cells
- Cloning
- Plasmid Extraction using Alkaline Lysis Method
- QuickChange Site Directed Mutagenesis
- Fusion PCR
- Golden Gate Cloning
Working with B. subtilis
Working with S. pneumoniae
Protein Methods
Assays
Team LMU-Munich
igem@bio.lmu.de
AG Thorsten MascherGroßhaderner Str. 2
82152 Planegg (Martinsried)
Hi there!
Welcome to our Wiki! I'm BaKillus, the pathogen-hunting microbe, and I'll guide you on this tour through our project. If you want to learn more about a specific step, you can simply close the tour and come back to it anytime you like. So let's start!
What's the problem?
First of all, what am I doing here? The problem is, pathogenic bacteria all around the world are becoming more and more resistant against antimicrobial drugs. One major reason for the trend is the inappropriate use of drugs. With my BaKillus super powers, I want to reduce this misuse and thus do my part to save global health.
Sensing of pathogens
To combat the pathogenic bacteria, I simply eavesdrop on their communication. Bacteria talk with each other via quorum sensing systems, which I use to detect them and trigger my responses.
Adhesion
The more specific and effective I can use my powers, the lower the danger is of provoking new resistance development. So I catch pathogens whenever I get hold of them and stick to them until my work is done.
Killing
Talking about my work - killing pathogens is finally what I am made for. In response to quorum sensing molecules of the pathogens, I export a range of antimicrobial substances leading to dissipation of biofilms and the killing of the targeted bacteria.
Suicide switch
When the job is done and all the bad guys are finished, you don't need a super hero anymore. So after fulfilling my work I say goodbye to the world by activating my suicide switch.
Application
Of course I'm not only a fictional hero, but a very real one. In two different prototypes, I could be used for diagnosis or treatment of pathogen-caused diseases. However, there is still a whole lot of regulational and economical questions that have to be answered before.
See you!
So now you know my short story - and it is time for me to return to my fight for a safer world. Feel free to take a closer look on my super powers, the process of my development or the plans for a medical application.
