Pfu: Difference between revisions
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(New page: ==Purpose== *Pfu is a thermostable DNA polymerase that can be used for PCR. ==Procurement== *You can buy Pfu for example at [[http://www.promega.com/catalog/catalogproducts.aspx?categoryn...) |
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==Procurement== | ==Procurement== | ||
*You can buy Pfu for example at | *You can buy Pfu for example at [http://www.promega.com/catalog/catalogproducts.aspx?categoryname=productleaf_1311&ckt=1 Promega] or [http://www.fermentas.com/catalog/modifyingenzymes/pfudnapolymer.htm Fermentas]. | ||
*NEB does for some reason not offer Pfu. | *NEB does for some reason not offer 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. | ||
*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== | |||
*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]] | |||
[[Category:Material]][[Category:Enzyme]][[Category:Polymerase]][[Category:DNA]] | [[Category:Material]][[Category:Enzyme]][[Category:Polymerase]][[Category:DNA]] | ||
Latest revision as of 15:11, 18 February 2009
Purpose
- Pfu is a thermostable DNA polymerase that can be used for PCR.
Procurement
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).
