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Star Republic: Guide for Biologists

PCR cleanup (SAP/ExoI)

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:

PCR products 10 ml
10X SAP buffer 1 ml
Shrimp Alkaline Phosphatase (SAP, 1 unit) 1 ml
E. coli exonuclease I (1 unit) 1 ml
ddH2O 7 ml

2. Incubate at 30 C for 30 minutes.

3. Inactivate the enzymes by heating at 80 C for 10-20 minutes.