ArticleJournal of assisted reproduction and genetics1996
Allele dropout in sequential PCR and FISH analysis of single cells (cell recycling).
Article in Journal of assisted reproduction and genetics, 1996. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- Clinical application of an Asian Screening Array-based preimplantation genetic testing workflow for various genetic disorders.Orphanet journal of rare diseases · 2026Article
- Haplotype Construction Using Embryos as Probands of the Pathogenic Variations in EXT1, CUL3, and HBA.Human mutation · 2026Article
- Prevention of lysosomal storage diseases and derivation of mutant stem cell lines by preimplantation genetic diagnosis.Molecular biology international · 2012Article
- Preimplantation genetic diagnosis (PGD) for nonsyndromic deafness by polar body and blastomere biopsy.Journal of assisted reproduction and genetics · 2009Article
- Preimplantation genetic diagnosis for ornithine transcarbamylase deficiency by simultaneous analysis of duplex-nested PCR and fluorescence in situ hybridization: a case report.Journal of Korean medical science · 2007Article
- Analysis of sex chromosomes in preimplantation genetic diagnosis for X-chromosome-linked disorders.Journal of assisted reproduction and genetics · 2002Article
- Molecular diagnostics in preimplantation genetic diagnosis.The Journal of molecular diagnostics : JMD · 2002Review
- 11q13 allelic imbalance discriminates pulmonary carcinoids from tumorlets. A microdissection-based genotyping approach useful in clinical practice.The American journal of pathology · 1999Article
- Preimplantation genetic diagnosis using fluorescent polymerase chain reaction: results and future developments.Journal of assisted reproduction and genetics · 1999Article
- Accuracy of preimplantation diagnosis of single-gene disorders by polar body analysis of oocytes.Journal of assisted reproduction and genetics · 1999Article
- Allele dropout in polar bodies and blastomeres.Journal of assisted reproduction and genetics · 1998Article
- Preimplantation diagnosis of thalassemias.Journal of assisted reproduction and genetics · 1998Article
- Preimplantation genetic diagnosis.Journal of assisted reproduction and genetics · 1996Article
- Review: preimplantation diagnosis of inherited disease.Journal of inherited metabolic disease · 1996Review
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
purposeOur purpose was to investigate the feasability of using sequential PCR and FISH analysis of single cells for preimplantation diagnosis.
methodsProtocols for sequential PCR and FISH analysis of a single fibroblast (cell recycling) were optimized for six loci and the rates of allele specific dropout (ADO) were determined.
resultsConditions that allow reliable genotyping of single cells in lysis buffer were not optimal for amplifying fibroblasts fixed to coverslips. After optimizing conditions, we observed a success rate of 85% for both analyses in sequential PCR-FISH experiments in single cells for the four loci studied. The individual success rates for each technique revealed a slightly higher rate for FISH (91-95%) than for PCR (85-87%) for single cells on coverslips. The presence of two hybridization signals in FISH experiments demonstrated that the failure to amplify both alleles from heterozygous cells on coverslips was due to true ADO, and not the loss of chromosomal material. The ADO rate observed on coverslips varied between 10 and 14%, which is significantly higher than that observed in solution, even after meticulous optimization.
conclusionsSequential PCR and FISH analysis of single cells remains an attractive possibility. However, until the problem of the increased rate of ADO is resolved, cell recycling should be applied to clinical preimplantation genetic analysis.
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