Pfu DNA Polymerase Mastermix [2X] is a premixed, ready-to-use solution containing Pfu DNA Polymerase, dNTPs, MgSO4 and Reaction Buffer at optimal concentrations for efficient amplification of DNA templates by PCR. To prepare the final PCR, only primers and template DNA are added. Pfu Mix contributes to highly reproducible PCR by reducing the risk of pipetting errors, miscalculation and contamination.
Pfu DNA polymerase, derived from the hyperthermophilic archae Pyrococcus furiosus, has been shown to exhibit superior thermostability and proofreading properties compared to other polymerases. Unlike Taq polymerase, highly thermostable Pfu DNA polymerase possesses 3' to 5' exonuclease proofreading activity that enables the polymerase to correct nucleotide-misincorporation errors. This means that Pfu DNA polymerase-generated PCR fragments will have fewer errors than Taq-generated PCR inserts. Using Pfu DNA polymerase in your PCR reactions results in blunt-ended PCR products, which are ideal for cloning into blunt-ended vectors. Use Pfu polymerase for techniques that require high-fidelity DNA synthesis.
UNIT DEFINITION
One unit (U) of Pfu polymerase is defined as the amount of enzyme needed to catalyze the incorporation of 10 nanomoles of deoxyribonucleotides into acid-insoluble material in 30 minutes at 70°C using herring sperm DNA as a substrate.
COMPOSITION OF THE Pfu MIX
Pfu DNA polymerase is supplied in 2X Pfu buffer, dNTPs, 3mM MgSO4 and bromophenol blue. Pfu mix buffer is a proprietary formulation optimized for robust performance in PCR.
Individual Pfu DNA Polymerase and Taq DNA Polymerase are available.
Features
- Convenient –Pfu DNA Polymerase in a ready-to-use Mix.
- High yields of PCR products with minimal optimization.
- Fast -saves time due to reduced number of pipetting steps.
- Reproducible -lower contamination and pipetting error risk.
Applications
- High fidelity PCR
- Routine PCR with high reproducibility
- Generation of PCR products for TA cloning
- Site-directed mutagenesis