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Human Reproduction Update, Vol.2, No.2 pp.137-152, 1996
© European Society of Human Reproduction and Embryology 1996; all rights reserved

Fluorescent polymerase chain reaction: Part I. A new method allowing genetic diagnosis and DNA fingerprinting of single cells

I Findlay0,1 and P Quirke2

0 Centre for Reproduction, Growth and Development, University of Leeds, Algernon Firth Building, Leeds General Infirmary, Leeds LS1 3EX, UK 2 Centre for Cancer Research and Molecular Pathology, Algernon Firth Building, Leeds General Infirmary, Leeds LS1 3EX, UK 1 Corresponding author

Abstract

The analysis of genetic material is fundamental to many medical and scientific applications. One method that has been used extensively for detection or diagnosis of genetic material, especially for single-gene defects or sex, is the polymerase chain reaction (PCR). It is particularly useful when the amount of genetic material is very limited, or samples have been degraded or fixed. In recent years, sensitivity has increased so that even single copies of genes within a single cell can be detected. This article discusses some of the difficulties involved with single-cell PCR and introduces single-cell fluorescent PCR and its potential applications. The use of fluorescent PCR to reduce misdiagnosis is discussed in detail.

Keywords: allelic dropout/DNA fingerprinting/fluorescent polymerase chain reaction/genetic diagnosis/preferential amplification


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