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Economical quantitative PCR (QPCR) reagent kits for use with all types of
fluorescent probes
Gothami Padmabandu Lisa Grismer Reinhold Mueller
Stratagene
Stratagenes Brilliant quantitative PCR and RT-PCR core reagent kits are compatible with many fluorescent detection technologies. Several versions have been created to optionally include dUTP/uracil-N-glycosylase (UNG) decontamination control reagents. All kits include SureStartTaq DNA polymerase , which provides the benefits of hot-start PCR, greatly reducing nonspecific primer annealing and extension that can lead to amplification of undesirable products. We evaluated the Brilliant quantitative PCR and RT-PCR core reagent kits in a variety of experiments, including the amplification of DNA and RNA targets followed by the detection of amplicon with various fluorescent chemistries. In this article we describe the data generated by Brilliant core reagents which include SureStart Taq and dUTP/UNG decontamination control.
Quantitative PCR has become the technology of choice for investigating gene
expression and gene copy number. One growing application is the validation of
differential expression signals seen with microarrays. While microarrays can
reveal expression level changes, the current level of noise in microarray
technology often warrants a secondary validation of expression level changes.
Similarly gene copy number measurements are subject to significant experimental
error. New quantitative PCR methods rely on real-time PCR analyses and generate
a fluorescence value that indicates the amount of product amplified to that
point in the reaction.1,2 In contrast to endpoint analysis, real-time
quantitation is based on measurements taken during the exponential phase of
amplification, a time when PCR efficiency is least influenced by reacti
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