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High Speed & Efficient PCR enzyme


KOD DNA polymerase exhibits an excellent elongation capability (approx. 130 bases/ sec) and strong 3'→5' exonuclease activity (proofreading activity). However, its sensitivity and efficiency are subject to interference from the enzyme or template concentration during PCR, due to its strong proofreading activity.

On the other hand, motif sequences of the exonuclease activity were identified by comparison of the sequences and 3-D structures among the family B DNA polymerases. By use of this information, the proofreading activity deficient <exo(-)> KOD DNA polymerase has been developed. This enzyme is useful for high speed, sensitive PCR.



KOD Dash (Code No. LDP-101) is a high speed, efficient PCR reagent based on Barns' method (1). This method uses a DNA polymerase mixture consisting of a DNA polymerase lacking a 3'→5' exonuclease (proofreading) activity (e.g. Taq DNA polymerase) and a small amount of an archaeal DNA polymerase with proofreading activity. Because the proofreading activity repairs mis-incorporated nucleotide bases causing the termination of a polymerase reaction, PCR with this mixed enzyme solution enables highly efficient amplification. In KOD Dash, the EXO(-) KOD (2) and KOD DNA polymerase are utilized.


This technology is utilized in Blend Taq (Code No. BTQ-101) and Blend Taq -Plus- (Code No.BTQ-201). These PCR reagents consist of Taq DNA polymerase and a small amount of an archaeal DNA polymerase with proofreading activity.

References

1)

W.M. Barns, Proc. Natl. Acad. Sci. USA, 91: 2216-2220 (1994)