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Nucleic Acids Research, 1990, Vol. 18, No. 4 973-978
© 1990

MOLECULAR BIOLOGY

The effect of replication errors on the mismatch analysis of PCR-amplified DNA

Jochen Reiss, Michael Krawczak, Manfred Schloesser, Michael Wagner and David N. Cooper

Institute of Human Genetics, University of Goettingen Gosslerstrasse 12 d, D-3400 Goettingen, FAG 1Molecular Genetics Section, Thrombosis Research Unit King's College Hospital School of Medicine, Denmark Hill, London SE5 8RS, UK

To whom correspondence should be addressed

Received September 25, 1989. Revised January 22, 1990. Accepted January 22, 1990.

The mismatch analysis of PCR-ampllfied DNA has generally assumed the absence of artificially introduced base substitutions in a significant proportion of the amplification product. This technique, however, differs from the direct sequencing of amplified DNA in that non-specific substitutions will render a molecule useless in analysis. The expected signal-to-noise ratio is heavily influenced by several parameters viz, initial template copy number, number of replication cycles, eventual product yield and the type of experimental system adopted. Mathematical modelling can be used to optimize fragment length with respect to the method applied and suggests as yet undescribed improvements such as partial modification or cleavage to optimize signal detection.


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