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Preimplantation diagnosis by whole-genome amplification, PCR amplification, and solid-phase minisequencing of blastomere DNA
1996 (English)In: Clinical Chemistry, ISSN 0009-9147, E-ISSN 1530-8561, Vol. 42, no 9, 1382-1390 p.Article in journal (Refereed) Published
Abstract [en]

We have developed a new method for preimplantation diagnosis of inherited diseases. Our procedure for the identification of point mutations in single cells combines whole-genome amplification using 15-mer random primers (primer extension preamplification, PEP) with a single locus-specific PCR amplification, followed by detection of the mutation by solid-phase minisequencing. The procedure was evaluated by detecting three disease-causing mutations and seven polymorphic nucleotides located on different human chromosomes from single granuloma and blastomere cells. The correct genotype of the cell was identified at 96% of the nucleotide positions analyzed, showing that a representative part of the genome is amplified during PEP. We estimate that PEP yielded at least 1000 copies of the genome. The quantitative nature of the solid-phase minisequencing method allowed us to notice that preferential amplification of one allele occurs at heterozygous loci during PEP, which is a potential problem in preimplantation diagnosis.

Place, publisher, year, edition, pages
1996. Vol. 42, no 9, 1382-1390 p.
National Category
Medical and Health Sciences
URN: urn:nbn:se:uu:diva-169821PubMedID: 8787693OAI: oai:DiVA.org:uu-169821DiVA: diva2:507786
Available from: 2012-03-06 Created: 2012-03-06 Last updated: 2012-04-10Bibliographically approved

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Syvänen, Ann-Christine
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