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PCR Product Clean-up

APPLICATION overview, CHALLENGES AND SOLUTION

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For Detailed Info Including:
  • Product overview
  • Performance data & figures
  • Specifications
  • Documents
  • FAQs
  • Ordering Info
  • Protocols
  • Publications

Application Overview

Shrimp Alkaline Phosphatase (SAP) and Exonuclease I (ExoI) have long been used together to prepare PCR products for subsequent sequencing or genotyping. These enzymes work synergistically to remove residual nucleotides and primers, which are common leftovers that can interfere with the accuracy of sequencing and genotyping assays.

See enzymes for

For Detailed Info Including:
  • Product overview
  • Performance data & figures
  • Specifications
  • Documents
  • FAQs
  • Ordering Info
  • Protocols
  • Publications

Application Overview

Shrimp Alkaline Phosphatase (SAP) and Exonuclease I (ExoI) have long been used together to prepare PCR products for subsequent sequencing or genotyping. These enzymes work synergistically to remove residual nucleotides and primers, which are common leftovers that can interfere with the accuracy of sequencing and genotyping assays.

THE PROBLEM These ENZYMEs SOLVE

Effective removal of residual nucleotides and primers is crucial for accurate sequencing and genotyping

Residual nucleotides and primers in PCR products can significantly interfere with downstream applications like sequencing or genotyping. These contaminants can affect the primer extension reaction, leading to inaccurate or unreliable results. Therefore, effective removal of these elements is crucial for the success of these sensitive techniques.

THE PROBLEM These ENZYME SOLVES

Effective removal of residual nucleotides and primers is crucial for accurate sequencing and genotyping

Residual nucleotides and primers in PCR products can significantly interfere with downstream applications like sequencing or genotyping. These contaminants can affect the primer extension reaction, leading to inaccurate or unreliable results. Therefore, effective removal of these elements is crucial for the success of these sensitive techniques.

Fig .

The Solution

SAP and ExoI offer a straightforward and effective solution to this problem. SAP dephosphorylates the residual nucleotides, rendering them ineffective, while ExoI degrades any remaining primers. By using these enzymes together, researchers can ensure that their PCR products are adequately purified, thereby enhancing the reliability and accuracy of subsequent sequencing or genotyping assays.

Protocol
Add to your PCR reactions after PCR is completed:

2 Units SAP
10 Units Exonuclease I
Incubate at 37°C for 15 min
Inactivate at 80°C for 15 min
The PCR product is now ready for sequencing or genotyping. The product may be stored at -20°C until required.

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Fig .  

Application Background