Technical Note: DNA Typing for Ovine MHC DRB1 Using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP)

The ovine major histocompatibilty complex (Ovar) class II DRB1 second exon was amplified by polymerase chain reaction (PCR) from DNA samples of 52 Suffolk sheep. Polymerase chain reaction products were characterized by the restriction fragment length polymorphism (RFLP) technique using nine restrict...

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Bibliographic Details
Published in:Journal of dairy science 2003-10, Vol.86 (10), p.3362-3365
Main Authors: Konnai, S, Nagaoka, Y, Takesima, S, Onuma, M, Aida, Y
Format: Article
Language:English
Subjects:
Alleles
animal genetics
Animal productions
Animals
Biological and medical sciences
DNA - analysis
DNA - chemistry
DNA Restriction Enzymes - metabolism
Food industries
Fundamental and applied biological sciences. Psychology
General aspects
genetic polymorphism
Genotype
immunology
major histocompatibility complex
Methods of analysis, processing and quality control, regulation, standards
Molecular Sequence Data
Ovar-DRB1 exon 2
PCR-RFLP
Polymerase Chain Reaction
Polymorphism, Restriction Fragment Length
rapid methods
restriction enzyme
restriction fragment length polymorphism
sheep
Sheep - genetics
Sheep - immunology
Suffolk (sheep breed)
Terrestrial animal productions
Vertebrates
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Summary:The ovine major histocompatibilty complex (Ovar) class II DRB1 second exon was amplified by polymerase chain reaction (PCR) from DNA samples of 52 Suffolk sheep. Polymerase chain reaction products were characterized by the restriction fragment length polymorphism (RFLP) technique using nine restriction enzymes, RsaI, HaeIII, SacI, SacII, DdeI, NciI, Hin1I, EcoRI, and BstNI, yielding 13 types. Sequencing of cloned PCR products identified 16 Ovar-DRB1 alleles. Collectively, all PCR-RFLP patterns exactly matched those predicted from DNA sequences. These findings strongly indicate that the PCR-RFLP method using a combination of nine restriction endonucleases is a very powerful tool in Ovar typing.
ISSN:0022-0302
1525-3198
DOI:10.3168/jds.s0022-0302(03)73939-3