LGC eBook - 2017 - 11

Accelerating Science Using Integrated PCR Tools * High-Multiplex Real-Time PCR Design

oligos result in discrete and predictable melting profiles for each target. As a result, multiple Catchers with unique Tm profiles can be
detected in the same reaction and in the same
color channel, significantly increasing the
number of targets that can be simultaneously
detected in the conventional four-channel
real-time thermocycler.
Another benefit of TOCE is the design versatility
of the Pitcher-Catcher detection system. Without
changing any other component in the assay, the
same target could be detected at different Tm
values simply by changing the sequence composition and/or length of the templating portion
of the Catcher. This controllable Tm property is a
key to the ease-of-design of a TOCE assay. Catcher
combinations are flexible, simplifying assay

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optimization through the selection of tagging
portion and Catcher combinations to give the
desired Tm profile. Moreover, these combinations
are interchangeable, so that tagging portions or
Catchers can easily be selected to minimize
nonspecific interactions.
TOCE is a major advancement in high-multiplex
real-time PCR capabilities, offering assay design
that greatly expands the capabilities of
conventional and next-generation, real-time
PCR instrumentation platforms. TOCE enables
high-multiplex assay design and development
across a wide range of applications, including
quantitative real-time PCR and highly selective
mutational analysis, providing a powerful new
tool for high-multiplex molecular assay
development in real time. n

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LGC eBook - 2017

Table of Contents for the Digital Edition of LGC eBook - 2017