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Title: Enhanced analysis of real-time PCR data by using a variable efficiency model: FPK-PCR
Authors: Lievens, Antoon ×
Van Aelst, Stefan
Van den Bulcke, M
Goetghebeur, E #
Issue Date: 2012
Publisher: OXFORD UNIV PRESS
Series Title: NUCLEIC ACIDS RESEARCH vol:40 issue:2 pages:e10-e10
Abstract: Current methodology in real-time Polymerase chain
reaction (PCR) analysis performs well provided PCR
efficiency remains constant over reactions. Yet,
small changes in efficiency can lead to large quantification errors. Particularly in biological samples,
the possible presence of inhibitors forms a challenge. We present a new approach to single
reaction efficiency calculation, called Full Process
Kinetics-PCR (FPK-PCR). It combines a kinetically
more realistic model with flexible adaptation to the
full range of data. By reconstructing the entire chain
of cycle efficiencies, rather than restricting the
focus on a ‘window of application’, one extracts
additional information and loses a level of arbitrariness. The maximal efficiency estimates returned
by the model are comparable in accuracy and
precision to both the golden standard of serial
dilution and other single reaction efficiency
methods. The cycle-to-cycle changes in efficiency,
as described by the FPK-PCR procedure, stay considerably closer to the data than those from other
S-shaped models. The assessment of individual
cycle efficiencies returns more information than
other single efficiency methods. It allows in-depth
interpretation of real-time PCR data and reconstruction of the fluorescence data, providing quality
control. Finally, by implementing a global efficiency
model, reproducibility is improved as the selection
of a window of application is avoided.
ISSN: 0305-1048
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Non-KU Leuven Association publications
× corresponding author
# (joint) last author

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