title: Haplotyping the human leukocyte antigen system from single chromosomes.
abstract: We describe a method for determining the parental HLA haplotypes of a single individual without recourse to conventional segregation genetics. Blood samples were cultured to identify and sort chromosome 6 by bivariate flow cytometry. Single chromosome 6 amplification products were confirmed with a single nucleotide polymorphism (SNP) array and verified by deep sequencing to enable assignment of both alleles at the HLA loci, defining the two haplotypes. This study exemplifies a rapid and efficient method of haplotyping that can be applied to any chromosome pair, or indeed all chromosome pairs, using a single sorting operation. The method represents a cost-effective approach to complete phasing of SNPs, which will facilitate a deeper understanding of the links between SNPs, gene regulation and protein function.
authors.affiliation: Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, Australia.|Preimplantation Genetic Diagnosis, Melbourne IVF, Melbourne, VIC, Australia.
authors.forename: Nicholas M
authors.initials: NM
authors.lastname: Murphy
authors.affiliation: Flow Cytometry &Imaging, Murdoch Children's Research Institute, Parkville, VIC, Australia.
authors.forename: Matthew
authors.initials: M
authors.lastname: Burton
authors.affiliation: Bioinformatics Platform, Monash University, Clayton, VIC, Australia.
authors.forename: David R
authors.initials: DR
authors.lastname: Powell
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC, Australia.|Australian Regenerative Medicine Institute, Monash University, Clayton, VIC, Australia.
authors.forename: Fernando J
authors.initials: FJ
authors.lastname: Rossello
authors.affiliation: Institute for Immunology and Infectious Diseases, Murdoch University, Murdoch, WA, Australia.
authors.forename: Don
authors.initials: D
authors.lastname: Cooper
authors.affiliation: Institute for Immunology and Infectious Diseases, Murdoch University, Murdoch, WA, Australia.
authors.forename: Abha
authors.initials: A
authors.lastname: Chopra
authors.affiliation: Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, Australia.
authors.forename: Ming Je
authors.initials: MJ
authors.lastname: Hsieh
authors.affiliation: Conexio Genomics, Fremantle, WA, Australia.
authors.forename: David C
authors.initials: DC
authors.lastname: Sayer
authors.affiliation: Australian Genome Research Facility, The Walter and Eliza Hall Institute of Medical Research, Royal Parade, Parkville, VIC, Australia.
authors.forename: Lavinia
authors.initials: L
authors.lastname: Gordon
authors.affiliation: Victorian Clinical Genetics Services, Parkville, VIC, Australia.|Department of Paediatrics, University of Melbourne, Parkville, VIC, Australia.
authors.forename: Mark D
authors.initials: MD
authors.lastname: Pertile
authors.affiliation: Transplant Services, Australian Red Cross Blood Service, Parkville, VIC, Australia.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Tait
authors.affiliation: Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, Australia.
authors.forename: Helen R
authors.initials: HR
authors.lastname: Irving
authors.affiliation: Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, Australia.
authors.forename: Colin W
authors.initials: CW
authors.lastname: Pouton
mesh_terms: D002896:Chromosomes, Human, Pair 6 / Q000235:genetics*; D020732:Cytogenetic Analysis / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D060005:Genotyping Techniques / Q000379:methods*; D006680:HLA Antigens / Q000235:genetics*; D006239:Haplotypes; D006801:Humans; D020641:Polymorphism, Single Nucleotide
chemical_list: D006680:HLA Antigens