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E. coli GFP transformation: Difference between revisions
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''Timescale:'' | ''Timescale:'' | ||
* Day 1 - prepare bacterial culture plates, incubate at room temperature in the dark | * Day 1 - prepare bacterial culture plates (1 per 4 people), incubate at room temperature (18-25C) in the dark for 3-4 days (or incubate at 37C for 18-24 hours, but results from this method not as good). | ||
* Before session - make agar plates and refrigerate | * Before session - make agar plates and refrigerate (these can be left for up to 7 days). Make transformation buffer. Dispense plasmid DNA into tubes. Prepare recovery broth. | ||
* Day 5 - | * Day 5 - Transformation. Warm plates to 37C. | ||
* Incubate plates overnight | * Incubate plates overnight at 37C. | ||
* Day 6 - Examine results | * Day 6 - Examine results (or store plates in fridge until viewing time) | ||
* Clean up - destroy cultures in autoclave. Disinfect all working materials. | |||
* | |||
''Storage when materials arrive:'' | ''Storage when materials arrive:'' | ||
* | * Plasmids must be stored at -20. | ||
* E. coli should be stored at 4C | |||
* Once E. coli pack is opened needs to be used in a few days. | * Once E. coli pack is opened needs to be used in a few days. | ||
* Shipped first class post mon - thurs | * Shipped first class post mon - thurs | ||
''Anything we need that we don't already have and doesn't come with the kit?'' | |||
* We'll need a fridge to store E. coli culture on arrival, LB plates once made, and culture plates if there's a delay between incubation and visualisation. | |||
* 37C incubator (investigate using environmental chamber) | |||
* 42C water bath (look at using new hot plate, check size sufficient) | |||
* Disinfectant (We currently have bleach and IPA) | |||
== Legal == | == Legal == | ||
