E. coli GFP transformation: Difference between revisions
From London Hackspace Wiki
|
|
| (48 intermediate revisions by 4 users not shown) |
| Line 1: |
Line 1: |
| = 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
| |
|
| |
| [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.
| |
|
| |
| == 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]]
| |
Latest revision as of 18:26, 2 April 2015
