Pfu

Purpose

• Pfu is a thermostable DNA polymerase that can be used for PCR.

Procurement

• You can buy Pfu for example at Promega or Fermentas.
• NEB does for some reason not offer Pfu.

Use

• Pfu has proof reading activity which makes it more accurate than for example Taq.
• Pfu does not exhibit 5' -> 3' exonuclease activity, which prevents degradation of the primers in mutagenesis methods based on ligation of primers or elongated primers.
• Pfu is slower than Taq and one should calculate 2 min per 1000 bp (1 min for Taq) (see Promega Pfu description).

Characteristics

• Elogation speed (bp/sec) tends to be lower than for example Taq.
• Build-in "Hotstart", because it's only active at high temperature, unlike Taq which has some activity at lower temperatures too
• Can degrade oligonucleotides, causing smearing (on gel).

Links

Smolke:Pfu Buffer