Artificial insemination of single women and lesbian women with donor semen: Recycling the single cell to detect specific chromosomes and to investigate specific gene sequences

We have developed a new procedure, called cell recycling, whichcombines the two powerful techniques of polymerase chain reaction(PCR) and fluorescent in-situ hybridization (FISH) on the samesingle cell. A fixed cell is used as the DNA template for PCR,prior to the FISH analysis. Using single blastomeres from mouseembryos as a model system, cell recycling procedures detectthe single-copy -haemoglobin gene sequence at an efficiencyof 70% as well as sex chromosome constitution at an efficiencyof 74% in the same single cell. Cell recycling will increasethe success rate of pregnancy following preimplantation diagnosisfor a specific gene defect by identifying embryos with chromosomalabnormalities and eliminating them from the transfer procedure.