PureIT ExoZAP PCR CleanUp consists of a balanced combination of a heat labile Exonuclease (HL-ExoI) I and a recombinant Shrimp Alkaline Phosphatase (rSAP).

Treatment of amplified PCR products with PureIT ExoZAP PCR CleanUp, removes residual primers, single-stranded DNA and inactivates excess dNTPs by dephosphorylation. Add PureIT ExoZAP PCR CleanUp directly to the reaction containing the amplified PCR product. After treatment at 37 °C for minimum 2 minutes, PureIT ExoZAP PCR CleanUp is completely inactivated by heating at 80 °C for minimum 3 minutes. 

PROTOCOL

This protocol serves as a guideline for clean-up of 5 μl PCR product using PureIT ExoZAP PCR CleanUp*.

• Take out PureIT ExoZAP PCR CleanUp from the -20 °C freezer.
• Keep PureIT ExoZAP PCR CleanUp on ice at all times.
• Add 2 μl PureIT ExoZAP PCR CleanUp** to 5 μl of amplified PCR product.
• Mix well and spin down.
• Incubate the reaction at 37 °C for 2 – 5 minutes to degrade remaining primers, single-stranded DNA and to inactivate excess nucleotides by dephosphorylation.
• Incubate at 80 °C to completely inactivate PureIT ExoZAP PCR CleanUp for 3 – 10 minutes.
• The cleaned up PCR product can now be used for downstream applications such DNA sequencing, primer extension experiment or SNP analysis.
• After treatment the PCR products can be stored at -20 °C

*If treating PCR product of higher volume, then increase proportionally the amount of PureIT ExoZAP PCR CleanUp.

**PureIT ExoZAP PCR CleanUp works in PCR buffer