PCR product clean-up
SAP has for years been utilized, together with Exonuclease I, to offer a simple and effective method for removing nucleotides and primers from PCR products prior to sequencing or genotyping.
In principle, SAP dephosphorylates nucleotides and ExoI degrades primers which would otherwise interfere with the primer extension reaction.
Protocol
Add to your PCR reactions after PCR is completed:
- 2 Units SAP
- 10 Units Exonuclease I
- Incubate at 37°C for 15 min
- Inactivate at 80°C for 15 min
The PCR product is now ready for sequencing or genotyping. The product may be stored at -20°C until required.
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