A new method for typing bovine major histocompatibility complex class II DRB3 alleles by combining two established PCR sequence-based techniques

• Published in: Tissue antigens Vol. 78; no. 3; pp. 208 - 213
• Main Authors: Takeshima, S.-N., Matsumoto, Y., Miyasaka, T., Arainga-Ramirez, M., Saito, H., Onuma, M., Aida, Y.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01-09-2011
• Subjects: allele genotyping; Animals; Base Sequence; bovine leukocyte antigen (BoLA) class II DRB3 genes; Cattle; Computer programs; DNA - genetics; DNA Primers - chemistry; DNA Primers - genetics; Genotype; Genotyping; Heterozygote; Histocompatibility Antigens Class II - genetics; Leukocytes; Major histocompatibility complex; Molecular Sequence Data; Polymerase chain reaction; Polymerase Chain Reaction - methods; polymerase chain reaction sequence-based typing; Primers; Sequence Analysis, DNA; software; Tissue typing
• ISSN: 0001-2815; 1399-0039
• DOI: 10.1111/j.1399-0039.2011.01708.x

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.