title: Recycling the single cell to detect specific chromosomes and to investigate specific gene sequences.
abstract: We have developed a new procedure, called cell recycling, which combines the two powerful techniques of polymerase chain reaction (PCR) and fluorescent in-situ hybridization (FISH) on the same single cell. A fixed cell is used as the DNA template for PCR, prior to the FISH analysis. Using single blastomeres from mouse embryos as a model system, cell recycling procedures detect the single-copy beta-haemoglobin gene sequence at an efficiency of 70% as well as sex chromosome constitution at an efficiency of 74% in the same single cell. Cell recycling will increase the success rate of pregnancy following preimplantation diagnosis for a specific gene defect by identifying embryos with chromosomal abnormalities and eliminating them from the transfer procedure.
authors.affiliation: Molecular Embryology Unit, Institute of Child Health, London, UK.
authors.forename: A
authors.initials: A
authors.lastname: Thornhill
authors.forename: C
authors.initials: C
authors.lastname: Holding
authors.forename: M
authors.initials: M
authors.lastname: Monk
mesh_terms: D000818:Animals; D001757:Blastomeres / Q000648:ultrastructure*; D002875:Chromosomes / Q000648:ultrastructure*; D015342:DNA Probes; D005260:Female; D005914:Globins / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D016133:Polymerase Chain Reaction; D011247:Pregnancy
chemical_list: D015342:DNA Probes; D005914:Globins