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J Biomol Tech. 2007 February; 18(1): 44–45.
PMCID: PMC2291905

P128-M SYBR Premix Ex Taq (Perfect Real Time): An Investigation of Instrument Compatibility

J. Gibson1 and T. Bio Inc.2

Abstract

In the early 1990s the discovery that Ethidium bromide (EtBr) could be used to monitor PCR cycles through binding double stranded DNA (dsDNA), resulting in a fluorescence increase, ushered in a new age in PCR. By the end of the decade new detection methods using probes (i.e. TaqMan) and instrumentation supported the launch of quantitative real time PCR (qPCR) technology. Since then many instruments and detection chemistries (FRET Hybrids, Molecular Beacons, SYBR Green and others) have been developed. Among these chemistries, the intercalating chemical SYBR Green is the most similar to EtBr and widely used. It is a fluorophore that binds dsDNA in the minor groove in a non-sequence-dependent manor. Upon binding, its fluorescence increases to ~10 times that of EtBr. This detection method is simple, sensitive and relatively inexpensive to other methods currently available. Since SYBR Green detects all dsDNA, primer dimers form during PCR will also be detected and quantified. These primer dimers and non-specific amplification can be minimized by using a “hot start” polymerase which prevents reaction initiation. To help alleviate detection of primer-dimers and other non specific amplification Takara has created SYBR Premix Ex Taq (Perfect Real Time). A convenient premix consisting of Takara’s high sensitivity-high performance Ex Taq Hot Start DNA Polymerase and SYBR Green I. Here we investigate this SYBR Green/ Ex Taq Hot Start combination and its compatibility with several qPCR instruments.


Articles from Journal of Biomolecular Techniques : JBT are provided here courtesy of The Association of Biomolecular Resource Facilities