Published in:
Open Access
01-05-2007 | INVITED EDITORIAL
Pitfalls in the normalization of real-time polymerase chain reaction data
Authors:
M. C. Hendriks-Balk, Prof. M. C. Michel, A. E. Alewijnse
Published in:
Basic Research in Cardiology
|
Issue 3/2007
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Abstract
Real-time polymerase chain reaction (PCR) is commonly used
for a sensitive and specific quantification of messenger RNA (mRNA). The
levels of mRNA are frequently compared between two or more experimental
groups. However, such comparisons require normalization procedures,
and reference genes are frequently used for this purpose. We discuss pitfalls
in normalization and specifically in the choice of reference genes. Reference
genes, which prove suitable for some experimental conditions, are not necessarily
similarly appropriate for others. Therefore,a proper validation of the
suitability of a given reference gene or sets thereof is required for each experimental
setting. Several computer programmes are available to aid such
validation.