PCR Clean-Up

PureIT ExoZAP

One-step PCR Clean-up for optimal sequencing results. Clean-up results in just 15 minutes.


FEATURES

  • One-step PCR product clean up
  • Fast 15 min protocol
  • No need for spin columns or magnetic beads
  • Treatment improves downstream applications such as DNA sequencing and SNP analysis
  • Minimal loss of PCR products during treatment


DESCRIPTION

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 5 minutes PureIT ExoZAP PCR CleanUp is completely inactivated by heating at 80 °C for 10 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 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.
  • 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 buffers.

PureIT ExoZAP PCR CleanUp IMPROVES QUALITY OF DNA SEQUENCING  

PCR product of 866 bp was amplified using TEMPase DNA Polymerase in Ammonium Buffer. The PCR product was spiked with dNTP’s and primers and then (A) treated without and (B) with PureIT ExoZAP PCR CleanUp prior to Sanger sequencing. All samples were analyzed in triplicates. Sanger sequencing result was compared by graphically plotting the number of base calls with certain quality values (Phred score). Treatment of PCR products with PureIT ExoZAP significantly improves the quality of DNA sequencing.


COMPARISON OF PureIT ExoZAP PCR CleanUp WITH TWO COMPETITORS  

Treatment of a 866 bp PCR product spiked with dNTP’s and primers by PureIT ExoZAP PCR CleanUp PCR, was compared to treatment by equivalent products from two well-recognised competitors, A and B. All samples were analyzed in triplicates. Sanger sequencing result was compared by graphically plotting the number of base calls with certain quality values (Phred score). PureIT ExoZAP treatment showed results equivalent to the two market leading competitors, A and B. Furthermore, the results for PureIT ExoZAP is obtained in just 15 min (5 min + 10 min) compared to protocols from competitor I and II of 30 min (15 min + 15 min).

Product info

PureIT ExoZAP

Product number

 

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