E. coli GFP transformation

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. 40C bath.

Proper storage.

Centrifuge - bigger : hold

Miniprep kit

Maxiprep kit

Self-cloning

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 The Genetically Modified Organisms (Contained Use) Regulations 2000 (PDF)

Here is a summary of those regulations: GMO Regulations 2000 summary