Source: the Pfu-DNA-Polymerase is a thermostabile polymerase of ca. 92 kDa in size, original isolated from the hyperthermophile archaebacteria Pyrococcus Furiosus. The enzyme replicates DNA at an optimum of 75°C, by catalysing polymerisation of nucleotides to duplex-DNA in 5’ -> 3’-direction in the presence of magnesium and privileged by MgSO4.

Different to Taq-DNA-Polymerase, the Pfu-DNA Polymerase additionally has a 3’ -> 5’-exonuklease-activity, the so called „proofreading-activity“, which enables the correction of wrong build-in nucleotides and, therefore, the establishment of almost faultless PCR-products. The precision of the DNA-synthesis is in contrast to the Taq-DNA-Polymerase about 12 times as much higher, the error rate is about 0,2 x 10 -5.

The PCR products produced by the Pfu-DNA-Polymerase have „blunt ends“ and can directly be used in ligation reactions. The extension rate of the Pfu-DNA-Polymerase is about 0,5 kb/min, therefore, 1 - 2 min extension time/kb fragment length is recommended.

Field of applications: PCR- und Primer-Extensions reactions, which need a high precision.

Quality control: use in test PCRs; examination on DNA-background.