Pfu

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*Pfu has proof reading activity which makes it more accurate than for example Taq.
*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 does not exhibit 5' -> 3' exonuclease activity, which prevents degradation of the primers in mutagenesis methods based on ligation of primers or elongated primers.
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*Pfu is slower than Taq and one should calculate 2 min per 1000 bp (1 min for Taq) ([http://www.promega.com/tbs/9pim774/9pim774.html see Promega Pfu description]).
==Characteristics==
==Characteristics==
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*Build-in "Hotstart", because it's only active at high temperature, unlike Taq which has some activity at lower temperatures too
*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).  
*Can degrade oligonucleotides, causing smearing (on gel).  
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==Links==
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[[Smolke:Pfu Buffer]]
[[Category:Material]][[Category:Enzyme]][[Category:Polymerase]][[Category:DNA]]
[[Category:Material]][[Category:Enzyme]][[Category:Polymerase]][[Category:DNA]]

Current revision

Contents

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

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