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Antibodies for Hot Start PCR


The thermostable DNA polymerases show a low background activity at room temperature during the preparation of a PCR reaction solution. The activity of the DNA polymerases (i.e. polymerase or 3'→5' exonuclease activity) at room temperature causes the generation of primer dimers or non-specific amplification.



Fig.1 Generation of primer dimers

Fig. 2 Degradation of primers
(Archaeal DNA polymerase)



Neutralization antibodies for the polymerase or proofreading activity are effective to inhibit the reactions of DNA polymerases below 60ºC. The antibodies are inactivated at the first denaturation step in the PCR reaction (Hot start PCR).


Antibodies against the polymerase and 3'→5' exonuclease activities of KOD DNA polymerase have been developed (1). These antibodies are used in KOD -Plus- (Code No. KOD-201) and KOD FX (Code No. KFX-101).


Antibodies against the polymerase activity of Taq/Tth DNA polymerases have also been developed. These antibodies are used in Realtime PCR Master Mix series (QPK-101, 201, 212), RNA-direct Realtime PCR Master Mix (QRT-101, 201), and RT-PCR Quick Master Mix (PCR-311).


The anti-Taq DNA polymerase antibody is available as "anti-Taq high (Code No. TCP-101).

Fig. 3 Principle of hot start PCR

References

1)

Mizuguchi H, Nakatsuji M, Fujiwara S, Takagi M and Imanaka T, J Biochem.,126: 762-8 (1999)