NeoProof DNA polymerase


  • NeoProof DNA polymerase NB-60-0006 : a recombinant enzyme with high fidelity and excellent PCR performance.
  • Includes 3´→5´ exonuclease proofreading for improved accuracy.
  • Efficiently amplifies longer PCR products (≤10 kb) and enables site-directed mutagenesis.
  • Recommended for routine cloning.
  • Error rate comparable to Pfu and Kod DNA polymerases, but significantly lower than Taq DNA polymerases.


Product Components and Storage

  • Supplied with a 10× Reaction Buffer and 5× Stabilizer Solution
  • Recommended storage at -20 °C in a constant temperature freezer
  • Stability until the expiry date if stored as specified
  • Unit definition: incorporation of 10 nmoles of dNTPs in 30 minutes at 72 °C


PCR Protocol

  • Standard protocol provided as a general guideline for PCR amplification
  • Guidelines for reaction setup, including component order and assembly on ice
  • Cycling parameters for denaturation, annealing, and extension
  • Gel electrophoresis for analyzing PCR products


PCR Design Considerations

  • Designing PCR primers with appropriate length (15-30 bases)
  • Recommendations for primer sequences to improve PCR yield and avoid degradation
  • Guidelines for primer GC content and avoidance of internal secondary structure
  • Recommendations to prevent primer-dimer formation and non-specific primer annealing
  • Importance of similar melting temperatures (Tm) for both primers


DNA Template

  • Recommended amounts of starting material based on quality and complexity
  • Guidelines for using genomic DNA templates and cDNA synthesis reaction templates
  • Caution against exceeding 10% of the final PCR reaction volume for cDNA templates


Enzyme Concentration

  • Recommended enzyme concentration of 1.25 U (0.5 μL) in a 50 μL reaction
  • Higher enzyme volumes (up to 2.5 U) for amplifying abundant templates (>50 ng gDNA)
  • Dilution options for convenience during PCR assembly


Quality Control Assays

  • Purity evaluation through SDS-PAGE and Coomassie Blue staining
  • Evaluation of genomic DNA contamination through PCR
  • Nuclease assays to check for nicking or cutting of nucleic acids
  • Functional assay using different-sized DNA fragments from human genomic DNA