AS ONE International’s PureAsOne PCR CleanUp Kit consists of heat labile Exonuclease I (Enz A) and a recombinant Shrimp Alkaline Phosphatase (Enz B). Treatment of PCR products with AS ONE PCR CleanUp Kit removes residual primers and inactivates dNTPs.
With PureAsOne PCR CleanUp Kit there is no need for time-consuming gel, column or magnetic bead purifications. And both short and long stranded PCR products are left completely intact.
– Easy handling
– Improved DNA sequencing results
– Time saving
– Fast – 5 minutes incubation time only
– Minimal loss of PCR products
Simply add the two reagents from the PureAsOne PCR CleanUp Kit to the reaction containing the amplified PCR product. Enz A degrades DNA primers, while Enz B inactivates dNTPs by dephosphorylation. After treatment at 37 °C for 2-5 minutes, both enzymes are completely inactivated by heating at 80 °C for 3-10 minutes.
Treated samples are ready for downstream applications such as DNA Sequencing and SNP analysis.
Fig 1. Comparison of Sanger sequencing results without and with the treatment of PureAsOne PCR CleanUp Kit. PCR product of 866 bp was amplified using Hot Start DNA Polymerase in Ammonium Buffer. The PCR product was spiked with dNTP’s and primers and then (A) treated without and (B) with PureAsOne PCR CleanUp Kit 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 PureAsOne significantly improves the quality of DNA sequencing.