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From 2008.igem.org

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Revision as of 09:43, 11 June 2008

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The protocols we used

• LB medium preparation
• Plasmid resuspension from IGEM paper spots
• Transformation
• Plasmid extraction
• BioBrick digestion with restriction enzymes
• DNA gel extraction
• Antarctic Phosphatase
• Ligation
• PCR

Antarctic Phosphatase

(estimated time: 1 hour and 30 min)

Materials needed:

• NEB Antarctic Phosphatase
• 10X NEB Antarctic Phosphatase buffer
• Cut and gel-extracted vector

• Add the proper amount of 10X buffer to a final concentration of 1X (e.g. 2 µl of 10X buffer in a final volume of 20 µl).
• Add 1 µl of Antarctic Phosphatase (up to 5 µg of cut vector).
• Incubate at 37°C for 1 hour (Antarctic Phosphatase works).
• Incubate at 65°C for 15 min (Antarctic Phosphatase inactivation).

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