Thermal-stable polymerases: info

Andre Hamel hamel at ccu.umanitoba.ca
Thu Jun 17 17:40:17 EST 1993


In article <1993Jun17.125928.13820 at cc.umontreal.ca> coady at ERE.UMontreal.CA (Michael Coady) writes:
>In article <nash.183.0 at biologysx.lan.nrc.ca> nash at biologysx.lan.nrc.ca (John Nash) writes:
>>
>>I thought that Pfu was Deep Vent (or am I wrong)?
>>
>
>	This time you're wrong, John :-).  According to the Stratagene
>catalogue, Vent polymerase is derived from Thermococcus litoralis while
>Pfu is from Pyrococcus furiosus.  Both do seem to have similar polymerase,
>proofreading and thermostability characteristics, though.
>
>Mike Coady
>COADY at ERE.UMONTREAL.CA
>
>


John Nash was referring to the NEB DEEP Vent enzyme ... not Vent pol
                              ^^^^
DV pol comes from a Pyrococcus sp.. :-)

Personally I find NEB's DV exo- pol better than Stratagenes Pfu exo- pol.
                           ^^^                                  ^^^

Mind you I can't possibly exhaustively test out EVERY situation (buffers,
templates, primers, cycling conditions), but several side-by-side tests,
once again point to NEB thermostable pols as being the best. 

My past comparisons between Pfu and Vent were before either company came
out with exo- versions, and who knows, products tend to change with time
anyway. But based upon my sequencing results from using these DNA pols for pcr
cloning, I've found myself "leaning" towards the NEB Vent and DV versus
Pfu or Taq (Cetus' Taq seems best versus other vendors', regardless of ALL
manner of buffers used (storage, rx'n) .. but once again, based on limited
experience, who knows what subtle changes/differences may/may been
accountable for observed differences between products made by these enzymes
... whatever works, use it ... if it ain't broke, don't fix it?

regards, Andre
                            ********************
Andre Hamel                                 email: hamel at ccu.umanitoba.ca
Manitoba Veterinary Services                lab tel.: (204) 945-7630
Infectious & Genetic Disease                     FAX: (204) 945-8062     
Mol.Biol.Lab., Winnipeg, Manitoba, CANADA
                            ********************



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