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
Growing competent E. coli
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), LB - £66, 0.1M CaCl2 - £32.50, M9 salts - £57.60, MgSO4 - £20, bacteriological agar solution - £42.90, thiamine - ~£20, D glucose - ~£20, and plates. All links are only the first ones I found, maybe cheaper options available.
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
New lab
Costs
- Rent, lets say £500 a month
Options for new space
- Talk to Russ about accomodating us in the new space.
- Investigate funding from other sources as cannot fund an independent space ourselves.
Funding sources:
- Wellcome trust
- Arts catalyst
- Synthetic aesthetic
- UCL. Imperial.
- Workshops
- Kickstarter
- Businesses to get free kit from
- BBC for new publicity.
- NESTA
- NCBE
Requirements for lab
- Tiling / washable surfaces
- Sink
- Lockable
