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 (cannot find how to buy this yet), [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, [http://www.sigmaaldrich.com/catalog/product/SIGMA/M6030?lang=en&region=GB M9 salts] - £57.60, [http://www.sigmaaldrich.com/catalog/product/sigma/m3409?lang=en&region=GB MgSO4] - £20, [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 available.
-== 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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