E. coli GFP transformation: Difference between revisions
From London Hackspace Wiki
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| = Genetic modification =
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| We're interested in doing genetic modification on micro-organisms at the London Hackspace. Ideas for stuff to do:
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| == E coli transformed with PUC19 using ampicillin selection ==
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| === 28 mar 15 ===
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| * 2ml ecoli culture (bottle on shelf) incubated in 200ml LB (bottle in fridge)
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| * -ve control is untransformed ecoli from same culture put on LBA
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| * +ve control is transformed ecoli from same culture put on LB plates
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| * Transform ecoli with PUC19 using electroporation
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| * Plate onto LBA.
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| == GFP transformation ==
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| * The 'hello world' GFP experiment. Biorad do a [http://www.bio-rad.com/prd/en/UK/LSE/PDP/619b8f74-9d3f-4c2f-a795-8a27e67598b7/pGLO-Bacterial-Transformation-Kit kit]. Carolina do just the [http://www.carolina.com/biotechnology-plasmid-lambda-dna/pgreen-plasmid-1-ug-200-ul-0005-ugul/211449.pr?catId=&mCat=&sCat=&ssCat=&question=pgreen+plasmid plasmid]
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| == Growing competent E. coli ==
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| [http://wiki.london.hackspace.org.uk/w/images/a/af/DIYbio_Collaboration_protocols.pdf Here] is the protocol we used with UCL to grow competent cells during the making of the public biobrick. So we need to get E. coli ([http://www.bio-rad.com/prd/en/US/LSE/SKU/166-0408EDU/E.-coli-Strain-HB101-K-12 one possibility]. Could also try NCBE. They sell e. coli as a transformer kit replacement part. Finally these are suggested for schools by NCBE: Sciento, Blades Biological and Philip Harris Education), [http://www.sigmaaldrich.com/catalog/product/fluka/l3152?lang=en®ion=GB LB] - £66, [http://www.sigmaaldrich.com/catalog/product/sigma/21115?lang=en®ion=GB 0.1M CaCl2] - £32.50, [http://www.sigmaaldrich.com/catalog/product/SIGMA/M6030?lang=en®ion=GB M9 salts] - £57.60, [http://www.sigmaaldrich.com/catalog/product/sigma/m3409?lang=en®ion=GB MgSO4] - £20, [http://www.sigmaaldrich.com/catalog/product/sigma/l2025?lang=en®ion=GB bacteriological agar solution] - £42.90, [http://www.sigmaaldrich.com/catalog/product/sial/t4625?lang=en®ion=GB thiamine] - ~£20, [http://www.sigmaaldrich.com/catalog/product/sigma/g8769?lang=en®ion=GB D glucose] - ~£20, and plates. All links are only the first ones I found, maybe cheaper options available.
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| [[Category:Biohacking]]
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Latest revision as of 18:26, 2 April 2015
