General Guidelines for Long PCR Conditions and Enzyme Mixtures Efficient Long PCR results from the use of two polymerases: a
non-proofreading polymerase is the main polymerase in the reaction, and a
proofreading polymerase (3' to 5' exo) is present at a lower
concentration. Following the results of Cheng et al. (1) we have had success using Tth (ABI/Perkin-Elmer) as the main-component polymerase and Vent
(New England Biolabs) as the fractional-component polymerase. Other
combinations of proofreading and non-proofreading polymerases have been
used successfully for many applications. The buffer listed below works
well with Tth and Vent, but not with others. If you are interested in
using other polymerases make sure that you use compatible buffers. Error
rates for other polymerases can be found at http://research.nwfsc.noaa.gov/protocols/taq-errors.html
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