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Expand High Fidelity PCR System

For customers in the European Economic Area: Contains SVHC: octyl/nonylphenol ethoxylates. For further processing on its own or in a mixture as part of an IVD method and under controlled conditions only – acc. to Art. 56 (3) and 3 no. 23 REACH Regulation.

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Order information
Expand High Fidelity PCR System material number and pack size:
Material Number Pack Size
03310256103 custom fill
Will be supplied as "Expand High Fidelity". Unit of measure is "kU".
The enzyme is supplied without reaction buffer.
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Overview
Proofreading blend for accurate amplification of genomic DNA targets up to 5 kb using PCR.
Enzyme blend consisting of Taq DNA Polymerase and Tgo DNA Polymerase.
  • Improve fidelity of PCR.
    Use this enzyme blend with its threefold greater accuracy than Taq Polymerase for more precise amplification of longer DNA templates.
  • Maximize target yield.
    Minimize amplification of prematurely terminated products using an ideally formulated proofreading enzyme for increased full-length yields.
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Application
Use Expand High Fidelity PCR System for:
  • Routine amplification of DNA fragments up to 5 kb from all DNA
  • Amplification of DNA fragments up to 10 kb.
  • Labeling of PCR products with modified nucleotides (e.g., DIG-dUTP, biotin-dUTP, fluorescein-dUTP)
  • Manufacture of amplification mixtures for regulated applications (e.g., in vitro diagnostics, quality control), including validation
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Specification
Enzymes in Expand High Fidelity PCR System were originally isolated from the thermophilic eubacteria Thermus aquaticus (Taq) BM or Thermococcus gorgonarius (Tgo), both expressed in E. coli.
Enzyme acivities:
Taq Polymerase: Highly processive 5'-3' DNA polymerase; double-strand-specific 5'-3' exonuclease; no 3'-5' exonuclease activity
Tgo Polymerase: Highly processive 5'-3' DNA polymerase; double-strand-specific 3'-5' exonuclease (also known as proofreading activity); no 5'-3' exonuclease activity.
pH optimum: Approximately 8.9 (+20°C)
Temperature optimum:
Fragment length <3 kb: Approximately +72°C
Fragment length >3 kb: Approximately +68°C
Substrates: Incorporates dNTP, dUPT, various labeled or modified nucleotides (200 μmol/L each is recommended of normal dNTP, increased concentrations of variants)
Divalent ion requirement: Mg2+ (1.5 mmol/L standard concentration)
Recommended usage per 50 μL reaction: 2.5 U (0.7 μL)
Appearance: Clear, colorless solution
Storage buffer: Tris/HCl, 20 mmol/L; KCI, 100 mmol/L; EDTA, 0.1 mmol/L; DTT, 1 mmol/L; Nonidet P40, 0.5% (v/v); Tween 20, 0.5% (v/v); glycerol, 50% (v/v); pH approximately 8.0 at +4°C
Volume activity: ≥3.5 U/μL
RNases (MS2 RNA): Not detectable in up to 30 U after 1 hour incubation at +37°C.
Function test in PCR (200 ng human genomic DNA, 4.8 kb tPA fragment): Corresponds to specification
Stability: At -15 to -25°C within specification range for 24 months.