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Human Reproduction, Vol. 7, No. 5, pp. 630-636, 1992
© 1992 European Society of Human Reproduction and Embryology

other

Reliability of detection by polymerase chain reaction of the sickle cell-containing region of the {beta}-globin gene in single human blastomeres

Susan J. Pickering1,2,3, Josie M. McConnell1, Martin H. Johnson1,2 and Peter R. Braude2

1Department of Anatomy, University of Cambridge Downing Street, Cambridge CB2 3DY, UK 2Assisted Conception Research Unit, UMDS Department of Obstetrics and Gynaecology, St Thomas' Hospital London SE1 7EH, UK

Correspondence: 3To whom correspondence should be addressed at: Department of Anatomy, University of Cambridge, Downing Street, Cambridge CB2 3DY, UK

Human preimplantation embryos at various stages of development have been analysed using the polymerase chain reaction to amplify a 680 base pair fragment of the {beta}-globin gene. Successful amplification was achieved more frequently with DNA from intact embryos containing between one and 11 cells, single cumulus cells, oocytes which had failed to fertilize and polar bodies than from single blastomeres disaggregated from intact embryos and treated in an identical manner. The distribution of nuclei demonstrated using the nuclear chromophore diamino-phenyl-indole showed considerable inter-blastomere variation; however, no clear correlation between staining pattern and successful amplification was observed. The reason for the unreliable amplification of DNA from single blastomeres is unclear but this finding has important implications for preimplantation diagnosis of genetic disease.

Key words: PCR/{beta}-globin/human blastomere


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