title: A new method for typing bovine major histocompatibility complex class II DRB3 alleles by combining two established PCR sequence-based techniques.
abstract: Recently, two polymerase chain reaction sequence-based typing (PCR-SBT) methods were reported for the genotyping of the bovine leukocyte antigen (BoLA)-DRB3. One technique is a single PCR-SBT (sPCR-SBT) method that generates heterozygous sequences that are subsequently analyzed by the haplofinder program, while the other technique is a nested PCR-SBT (nPCR-SBT) method that allows the analysis of heterozygous sequences using the assign 400ATF software. In this study, these techniques were compared and then integrated to produce an improved genotyping method. The primer set used for sPCR-SBT was more accurate than those used for nPCR-SBT. Combining sPCR-SBT with the assign 400ATF software previously reported for nPCR-SBT enables rapid and accurate genotyping of a large number of DNA samples.
authors.affiliation: Viral Infectious Diseases Unit, RIKEN, Wako, Saitama, Japan.
authors.forename: S-N
authors.initials: SN
authors.lastname: Takeshima
authors.forename: Y
authors.initials: Y
authors.lastname: Matsumoto
authors.forename: T
authors.initials: T
authors.lastname: Miyasaka
authors.forename: M
authors.initials: M
authors.lastname: Arainga-Ramirez
authors.forename: H
authors.initials: H
authors.lastname: Saito
authors.forename: M
authors.initials: M
authors.lastname: Onuma
authors.forename: Y
authors.initials: Y
authors.lastname: Aida
mesh_terms: D000818:Animals; D001483:Base Sequence; D002417:Cattle; D004247:DNA / Q000235:genetics*; D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D005838:Genotype; D006579:Heterozygote; D000949:Histocompatibility Antigens Class II / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
chemical_list: C423282:BoLA-DRB3 antigen; D017931:DNA Primers; D000949:Histocompatibility Antigens Class II; D004247:DNA