PCR Clean-Up
Ampliqon has developed PureIT ExoZAP PCR CleanUp, which is an enzymatic PCR clean-up reagent preparing PCR products for various downstream applications such as sequencing, primer extension and SNP analysis.
PureIT ExoZAP PCR CleanUp
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 minimum 2 minutes, PureIT ExoZAP PCR CleanUp is 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*.
- 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 2 – 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 for 3 – 10 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 PureIT ExoZAP PCR CleanUp.
**PureIT ExoZAP PCR CleanUp works in PCR buffer
-One-step PCR product clean up
-Fast 5 min protocol
-No Sample Loss
-No need for spin columns or magnetic beads
-Treatment improves downstream applications such as DNA sequencing and SNP analysis
-Ideal for NGS library preparation protocols
