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Protocol for Dephosphorylation of DNA 5'-termini
(with FastAP™ Thermosensitive Alkaline Phosphatase)

1. Prepare the following reaction mixture:
   

   Linear DNA (~3 kb plasmid)	1 µg (~1 pmol termini)
   10X FastAP™ buffer	2 µl
   FastAP™ Thermosensitive Alkaline Phosphatase	1 µl (1 u)
   Water, nuclease-free	to 20 µl

2. Mix thoroughly, spin briefly and incubate at 37°C for 10 minutes.
3. Stop reaction by heating at 75°C for 5 minutes.

Note
For efficient dephosphorylation plasmid DNA should be free of RNA and genomic DNA.

Ordering Information

FastAP™ Thermosensitive Alkaline Phosphatase Water, nuclease-free

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Updated sausio 04, 2008 10:37