TaqPlus Precision PCR System
Thermostable DNA polymerases offer distinct advantages for specific PCR applications. Pfu DNA polymerase exhibits the lowest error rate of any thermostable DNA polymerase analyzed. This proofreading enzyme is also characterized by a relatively slow polymerization rate requiring the use of long extension times (i.e., ³2 minutes/kb of DNA target) to generate high yields of PCR product. Taq DNA polymerase exhibits a fivefold higher polymerization rate than Pfu DNA polymerase and is used routinely to generate high yields of PCR product £5 kb in length using short extension times (i.e., ~1 minute/kb of DNA target). Taq DNA polymerase possesses a significantly higher error rate than Pfu DNA polymerase. However, Taq2000 DNA polymerase is superior to other commercial Taq DNA polymerases in that this recombinant version of Taq DNA polymerase minimizes artifactual smearing in PCR amplification reactions requiring long extension times.
DNA polymerase mixtures, consisting of a non-proofreading DNA polymerase and a proofreading DNA polymerase, are typically employed to amplify longer DNA targets and to generate higher yields of PCR product than can be obtained using either DNA polymerase alone. Agilent offers two mixtures of Taq and Pfu DNA polymerases designed for PCR: the TaqPlus Long PCR system and the TaqPlus Precision PCR system. The TaqPlus Long PCR system typically generates the highest yields of PCR product and can be used to amplify unusually difficult or long targets (up to 35 kb). The TaqPlus Precision PCR system also generates relatively high yields of PCR product using short PCR extension times. The error rate, however, is significantly lower than the TaqPlus Long PCR system. In addition, the TaqPlus Precision PCR system successfully amplifies plasmid and lambda DNA templates up to 15 kb in length and single-copy genomic DNA templates up to 10 kb in length as well as templates that are difficult to amplify using single enzyme formulations.