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E. coli GFP transformation: Difference between revisions

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We're interested in doing genetic modification on micro-organisms at the London Hackspace. Ideas for stuff to do:
We're interested in doing genetic modification on micro-organisms at the London Hackspace. Ideas for stuff to do:
== GFP transformation ==


* The 'hello world' GFP experiment. Biorad do a [http://www.bio-rad.com/prd/en/UK/LSE/PDP/619b8f74-9d3f-4c2f-a795-8a27e67598b7/pGLO-Bacterial-Transformation-Kit kit]. Carolina do just the [http://www.carolina.com/biotechnology-plasmid-lambda-dna/pgreen-plasmid-1-ug-200-ul-0005-ugul/211449.pr?catId=&mCat=&sCat=&ssCat=&question=pgreen+plasmid plasmid]
* The 'hello world' GFP experiment. Biorad do a [http://www.bio-rad.com/prd/en/UK/LSE/PDP/619b8f74-9d3f-4c2f-a795-8a27e67598b7/pGLO-Bacterial-Transformation-Kit kit]. Carolina do just the [http://www.carolina.com/biotechnology-plasmid-lambda-dna/pgreen-plasmid-1-ug-200-ul-0005-ugul/211449.pr?catId=&mCat=&sCat=&ssCat=&question=pgreen+plasmid plasmid]
* Get some ideas from iGem teams
Needed for GFP experiment (that we don't already have):
Fridge?
Incubator.
Build or buy. Or could use hackspace environmental chamber. Can be programmed with right temp but non-sterile.
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.
=== Still need ===


Centrifuge
* GFP + antibiotic resistance plasmid.
- bigger : hold
* Ampicillin.
* Initial e. coli culture.
* Growth medium/ broth.


Miniprep kit
=== Already have ===
* Plates.
* Agar
* LB Broth
* Temp controlled water bath


Maxiprep kit


== Growing competent E. coli ==
== Growing competent E. coli ==
Mycoplasma
927

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