文献_whole genome amplification_WGA Methods express.pdfVIP

11-Genome-ch11-ppp 14/7/05 7:54 am Page 1 CHAPTER 11 Pre-implantation genetic diagnosis using whole genome amplification 1 2 3 Alan H. Handyside , Mark D. Robinson and Francesco Fiorentino 1London Bridge Fertility, Gynaecology and Genetics Centre, One St Thomas Street, London Bridge, London SE1 9RY, UK; 2School of Biology, University of Leeds, Leeds LS2 9JT, UK; 3School of Biology, University of Leeds, Leeds, UK; 3Laboratorio Genoma, Rome, Italy 1. INTRODUCTION Pre-implantation genetic diagnosis (PGD) following assisted conception is now well established clinically as an alternative to conventional pre-natal diagnosis in couples at risk of having children with an inherited disease (1). Controlled ovarian stimulation, egg collection by ultrasound-guided transvaginal needle aspiration and insemination with the partner’s washed sperm provide access to fertilized pre-implantation-stage embryos in vitro. Single cells, typically the first and second polar bodies and/or one or two blastomeres, are then removed by micromanipulation from each fertilized zygote or cleavage-stage embryo, respectively, for genetic analysis. This typically involves fluorescent in situ hybridization (FISH) and other molecular cytogenetic techniques for detection of