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Shrimp Alkaline Phosphatase (SAP) and E. coli exonuclease I (ExoI)
dephosphorylate dNTP and degrade single-stranded DNA (primer).
1. Make the following mix in a microcentrifuge tube:
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PCR products
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10 ml
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10X SAP buffer
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1 ml
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Shrimp Alkaline Phosphatase (SAP, 1 unit)
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1 ml
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E. coli exonuclease I (1 unit)
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1 ml
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ddH2O
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7 ml
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2. Incubate at 30 °C for 30 minutes.
3. Inactivate the enzymes by heating at 80 °C for 10-20 minutes.
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