PureIT ExoZAP PCR CleanUp

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

FEATURES

One-step PCR product clean up
Fast 5 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 866 bp PCR product was hereafter spiked with dNTP’s and primers and then (A) treated without or (B) with PureIT ExoZAP PCR CleanUp prior to Sanger Sequencing. All samples were analyzed in triplicates. Quality of Sanger sequencing was determined by analysis of base calls with high quality values (Phred score). Treatment of PCR products with PureIT ExoZAP PCR CleanUp significantly improves the quality of DNA sequencing.

COMPARISON OF PureIT ExoZAP PCR CleanUp WITH TWO COMPETITORS

Treatment by PureIT ExoZAP PCR CleanUp PCR, was compared to treatment by equivalent products from two well-recognised competitors A and B. PCR product of 866 bp was amplified using TEMPase DNA Polymerase in Ammonium Buffer. The 866 bp PCR product was hereafter spiked with dNTP’s and primers. All samples were analyzed in triplicates. Quality of the subsequent Sanger sequencing was determined by analysis of base calls with high quality values (Phred score). PureIT ExoZAP improves the quality of DNA Sequencing to the same extend as the two competitors A and B. Furthermore, the results for PureIT ExoZAP PCR CleanUp is obtained in just 15 min (5 + 10) compared to the recommended protocols from competitor A and B of 30 min (15 + 15)

Pricing Table Particle

PureIT ExoZAP PCR CleanUp

AL-Labortechnik e. U.