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From 2010.igem.org

MATERIALS
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- Heat block
- Spin

- Linear DNA
- 10X reaction buffer for AP used in reaction
- FastAP™ Thermosensitive Alkaline Phosphatase
- Nuclease-free water

- P10 micropipette

CHECK-LIST PROCEDURE
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- Prepare the following reaction mixture: Linear DNA (~3 kb plasmid) 1 μg (~1 pmol termini) 10X reaction buffer for AP used in reaction 2 μl FastAP™ Thermosensitive Alkaline Phosphatase 1 μl (1 u) Water, nuclease-free (#R0581) to 20 μl Total volume 20 μl
- Mix thoroughly, spin briefly and incubate 10 min at 37°C.
- Stop reaction by heating for 5 min at 75°C.

NOTES
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- For efficient dephosphorylation plasmid DNA should be free of RNA and genomic DNA.
- This protocol is suitable for removal of 3’ and 5’ -phosphate groups from DNA and RNA.