Multiplex polymerase chain reaction for sex determination of single mouse blastomeres

Using a multiplex nested polymerase chain reaction (PCR) method with single copy genes and a dinucleotide repeat locus for the mouse Y and X chromosomes respectively, it was possible to discriminate between single cells derived from male and female embryos. Using single cells, amplification of Sry a...

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Bibliographic Details
Published in:Human reproduction (Oxford) 1995-03, Vol.10 (3), p.743-748
Main Authors: DREESEN, J. C. F. M, DUMOULIN, J. C. M, EVERS, J. L. H, GERAEDTS, J. P. M, PIETERS, M. H. E. C
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Biological and medical sciences
Blastomeres
DNA Primers
Female
Fertilization in Vitro
Gynecology. Andrology. Obstetrics
Male
Management. Prenatal diagnosis
Medical sciences
Mice
Mice, Inbred C57BL
Mice, Inbred CBA
Molecular Sequence Data
Polymerase Chain Reaction
Pregnancy. Fetus. Placenta
Sex Determination Analysis - methods
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Summary:Using a multiplex nested polymerase chain reaction (PCR) method with single copy genes and a dinucleotide repeat locus for the mouse Y and X chromosomes respectively, it was possible to discriminate between single cells derived from male and female embryos. Using single cells, amplification of Sry and Zfy sequences was not evident in all cases. It could be calculated that, with the PCR method used, 0.04% [95% (confidence interval 0.00-2.03)] of the male embryos would erroneously be diagnosed as female if analysis is performed on two cells. The calculated chance for total amplification failure, if two cells are used for analysis, would be 1.4% [95% (confidence interval 0.04-8.04)]. The mouse embryo model proved to be a helpful tool to develop skills in the application of PCR for preimplantation genetic diagnosis at the single cell level.
ISSN:0268-1161
1460-2350
DOI:10.1093/oxfordjournals.humrep.a136025