Melbourne/Transformation Protocol shorter
From 2007.igem.org
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=====References===== | =====References===== | ||
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Revision as of 11:45, 28 September 2007
<Return to list of protocols> <Team home page>
- Applications:
- Amplification of Biobrick DNA for storage and use.
- Selection and amplification of ligated constructs.
- Time to complete protocol:
- Lab time: 10min, 10min, 10min, 15min.
- Waiting time: 45min, 15min, 1hour, overnight.
- Approximate cost of materials: $
Method from primary and secondary reagents
Primary & secondary Reagents Required including controls
- Competent cells (from -70degree freezer)
- DNA for transformation
- LB (cupboard)
- LB-agar plates with selective antibiotic (cool room)
Method including controls
- Add 1uL resuspended plasmid DNA to 50uL competent cells.
- Incubate on ice for 30min.
- Heat shock in water bath at 42 degrees for 1min.
- Incubate on ice for 10min.
- Add 1mL LB (flame tip before use).
- Incubate at 37 degrees for 30 minutes.
- Spin down cells and remove majority of LB.
- Resuspend cells in remaining LB.
- Under a bunsen spread resuspended bacteria on agar plate on selective antibiotic.
- Incubate plate overnight at 37 degrees.
- Place in cold room until needed.
Equipement Required
- 1.7ml microcentrifuge
- Ice box
- Pipettes
- 42 degree water bath (balance room)
- 37 degree incubator
- Bunsen burner
- Spreader
- ice
References
