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.
Expand High Fidelity PCR System material number and pack size: Material Number Pack Size 03310256103 custom fill
The enzyme is supplied without reaction buffer.
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.
- Improve fidelity of PCR.
- 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
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.
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)
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.