Difference between revisions of "E. coli GFP transformation"

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-= Genetic modification =
-We're interested in doing genetic modification on micro-organisms at the London Hackspace. Needed for GFP experiment (that we don't already have):
-Fridge?
-Incubator.
-	Build or buy.
-Cleaning:
-	Bleach, gloves, paper towels, hand cleaning gel.
-Bunsen burner + fuel.
-Glassware, pipettes, tools.
-Ethanol.
-	Lots.
-To apply for a license we will probably first need:
-Written protocols for stuff, especially cleaning up.
-Disposal.
-Chemical.
-Sterilisation.
-Do we need incineration? Find a waste disposal company that will take biohazard material?
-Restriction enzymes.
-	At least two.
-	Pricey?
-GFP plasmid.
-Antibiotics.
-	Ampicillin.
-Initial e. coli culture.
-Growth medium/ broth.
-	LB.
-Plates.
-	Standard 10cm plates.
-	Agarose.
-Ice.
-C bath.
-Proper storage.
-Centrifuge
-	- bigger : hold
-Miniprep kit
-Maxiprep kit
-== Growing competent E. coli ==
-[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.sigmaaldrich.com/catalog/product/fluka/l3152?lang=en&region=GB LB] - £66, [http://www.sigmaaldrich.com/catalog/product/sigma/21115?lang=en&region=GB 0.1M CaCl2] - £32.50, M9 salts, MgSO4, [http://www.sigmaaldrich.com/catalog/product/sigma/l2025?lang=en&region=GB bacteriological agar solution] - £42.90, [http://www.sigmaaldrich.com/catalog/product/sial/t4625?lang=en&region=GB thiamine] - ~£20, [http://www.sigmaaldrich.com/catalog/product/sigma/g8769?lang=en&region=GB D glucose] - ~£20, and plates. All links are only the first ones I found, maybe cheaper options.
-== Self-cloning ==
-[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.
-* plate spreaders
-* petri dishes
-* LB
-* Agar
-* Howard has a good protocol for transformation
-== Legal ==
-There are a number of things we have to do in order to set up a GM lab.
-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)
-Here is a summary of those regulations: [[Project:Biohacking/Genetic_modification/Regulations_2000_summary|GMO Regulations 2000 summary]]

Difference between revisions of "E. coli GFP transformation"

Latest revision as of 18:26, 2 April 2015

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