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Revision as of 06:52, 8 September 2009
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Protocols
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Transformation
(estimated time: 3 hours and 30 min + 12-16 hours overnight incubation)
- Put 1-2 µl of DNA resuspension or ligation into thaw TOP10 cells tube (every tube contains approximately 60 µl of competent cells).
- Incubate on ice for 30 min.
- Heat shock: 42°C for 1 min.
- Put transformed TOP10 tube on ice and then add 250 µl SOC medium.
- Incubate 1 hour at 37°C, 220 rpm.
- Plate 200 µl of the solution on a proper agar plate.
- Incubate overnight at 37°C.
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- LB agar plates with proper antibiotic
- Invitrogen TOP10 cells
- Resuspended DNA
- SOC medium
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