PureIT ExoZAP - PCR Clean-Up reagent

The PureIT ExoZAP - PCR Clean-Up reagent from Ampliqon can be used for enzymatic cleanup of your PCR products without the need for purification columns. With PureIT ExoZAP PCR products from less than 100bp to over 20kbp can be purified from primers and unincorporated dNTPs with no sample loss. The combination of HL-ExoI and a recombinant Shrimp Alkaline Phosphatase (rSAP) hydrolyzes excess primers and nucleotides in a single step within the PCR reaction tube!

ZAP-SAPexo purified samples are ready for use in downstream applications such as DNA sequencing or SNP analysis (single nucleotide polymorphism) after a simple 5 minute incubation.

Advantages:
• Fast. Only 5 minutes without centrifugation.
• Just one pipetting step for pure DNA sequencing samples!
• Simple enzymatic removal of excess primers and unincorporated nucleotides.
• Minimal loss of your PCR sample. Up to 100% recovery.

Our innovative PureIT ExoZAP solution offers a simple method to purify your PCR reactions from primers and unincorporated nucleotides. Only one pipetting step followed by a min. 2 minutes incubation at 37°C (digestion) and a min. 3 (-10) minutes incubation at 80°C (inactivation). One-step solution provides a simple way to enzymatically purify a PCR reaction. Just programme your PCR cycler after the PCR accordingly, pipet the PureIT ExoZAP reagent to each tube or well, start Cycler and you will have your purified PCR samples after 5 minutes ready for use in sequencing or SNP analyis.

After the enzymatic reaction with PureIT ExoZAP, primers and dNTPs will no longer interfere with DNA sequencing or SNP analysis, for example with our SNP Pol DNA polymerase.

Using PureIT ExoZAP offers the possibility to proceed with the sequencing reaction after the BigDye™ Cycle Sequencing reaction without the need for time consuming purification methods like the Centri-Sep columns from Princeton or the Genaxxon CentriPure purification plates.

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 treated (A) without PureIT ExoZAP PCR CleanUp (B) with PureIT ExoZAP PCR CleanUp
(2 +3 min) and (C) with an enzymatic PCR clean-up reagent from a competting brand (1+ 4 min) prior to Sanger sequencing.
All samples were analyzed in triplicates and according to manufactures recommendations. 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. Furthermore, PureIT ExoZAP PCR
CleanUp showed PCR clean-up results equivalent to the competing PCR clean-up reagent. *Data with Quality values less than
10 is not included.

Sanger sequencing results of the 866 bp PCR product. The depicted electropherograms are either after treatment with
PureIT ExoZAP PCR CleanUp (top) or untreated (bottom). Sequence shown is approximately the region from 250 - 300 bases.
Treatment of spiked PCR product with PureIT ExoZAP PCR CleanUp significantly improves the quality of Sanger Sequencing.
Treatment enhances signal intensity and eliminates background interference and base miscalls.