Pfu: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
(add link) |
||
| Line 15: | Line 15: | ||
*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). | ||
==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).
