E. coli GFP transformation: Difference between revisions
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| = Genetic modification =
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| We're interested in doing genetic modification on micro-organisms at the London Hackspace. Needed for GFP experiment (that we don't already have):
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| Fridge?
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| Incubator.
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| Build or buy.
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| Cleaning:
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| Bleach, gloves, paper towels, hand cleaning gel.
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| Bunsen burner + fuel.
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| Glassware, pipettes, tools.
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| Ethanol.
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| Lots.
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| To apply for a license we will probably first need:
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| Written protocols for stuff, especially cleaning up.
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| Disposal.
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| Chemical.
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| Sterilisation.
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| Do we need incineration? Find a waste disposal company that will take biohazard material?
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| Restriction enzymes.
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| At least two.
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| Pricey?
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| GFP plasmid.
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| Antibiotics.
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| Ampicillin.
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| Initial e. coli culture.
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| Growth medium/ broth.
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| LB.
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| Plates.
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| Standard 10cm plates.
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| Agarose.
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| Ice.
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| 40C bath.
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| Proper storage.
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| Centrifuge
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| - bigger : hold
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| Miniprep kit
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| Maxiprep kit
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| == Self-cloning ==
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| [http://www.ncbe.reading.ac.uk/ncbe/protocols/transformer.html Here's] a protocol to do a 'self-cloning' transformation, which we would not need a licence for, and so could do now to practise.
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| * plate spreaders
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| * petri dishes
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| * LB
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| * Agar
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| * Howard has a good protocol for transformation
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| == Legal ==
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| There are a number of things we have to do in order to set up a GM lab.
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| The relevant regulations are [http://www.legislation.gov.uk/uksi/2000/2831/pdfs/uksi_20002831_en.pdf The Genetically Modified Organisms (Contained Use) Regulations 2000] (PDF)
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| Here is a summary of those regulations: [[Project:Biohacking/Genetic_modification/Regulations_2000_summary|GMO Regulations 2000 summary]]
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Latest revision as of 18:26, 2 April 2015
