Nucleic Acids Research, Vol 26, Issue 15 3468-3472, Copyright © 1998 by Oxford University Press
AM Garvin, W Holzgreve and S Hahn
Single cell PCR is a powerful method for determining the genetic properties
of individual cells. In the instance of heterozygous loci, however,
preferential amplification of one allele can lead to allele drop out (ADO)
of the other allele. Fortunately ADO does not occur in all amplifications,
and is usually random when it does occur, with both alleles being equally
susceptible to drop out. Therefore pooling of results from multiple
independently amplified cells should greatly improve the analysis of
diallelic loci, and the misdiagnosis rate of diallelic loci should decrease
exponentially with the number of cells analysed. We have shown that this is
true and that multiplex PCR allows for the simultaneous identification of a
cell in a mixture of cells using microsatellite loci known to be
informative, and accurate genotyping at other loci. This approach has
practical applications to non-invasive prenatal diagnosis where small
numbers of fetal cells in the presence of maternal cells must be both
identified and genotyped at loci involved in genetic disease.
ARTICLES
Highly accurate analysis of heterozygous loci bysingle cell PCR
Department of Obstetrics and Gynaecology and Laboratory for Prenatal Medicine, University of Basel,46 Schanzenstrasse, Basel CH-4031, Switzerland.
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