PCR Clean-Up

PureIT ExoZAP Kit

PCR Clean-up for optimal sequencing results. Clean-up results in 5 minutes.


FEATURES

  • PCR product clean up
  • 5 min protocol
  • No need for spin columns or magnetic beads
  • Treatment improves downstream applications such as DNA sequencing and SNP analysis


DESCRIPTION

PureIT ExoZAP PCR CleanUp Kit consists 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 Kit, degrades residual primers, single-stranded DNA and inactivates excess dNTPs by dephosphorylation. Add PureIT ExoZAP PCR CleanUp Kit directly to the reaction containing the amplified PCR product. After treatment at 37 °C for minimum 2 minutes both enzymes are 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 Kit*.

 

  • Take out PureIT ExoZAP PCR CleanUp Kit from the -20 °C freezer.
  • Keep the components of PureIT ExoZAP PCR CleanUp Kit on ice at all times.
  • Add 1.5 μl rSAP and 0.5 μl HL-ExoI to 5 μl of amplified PCR product**.
  • Mix well and spin down.
  • Incubate the reaction at 37 °C for minimum 2 minutes to degrade remaining primers, single-stranded DNA and to inactivate excess nucleotides by dephosphorylation.
  • Incubate at 80 °C to completely inactivate both enzymes for minimum 3 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 HL-ExoI and rSAP.

**The enzymes within PureIT ExoZAP PCR CleanUp Kit 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.

Product info

PureIT ExoZAP Kit

Product number
HL-ExoI
rSAP

 

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