A rapid method for genotyping mice for T cell receptor V beta a and V beta b haplotypes by PCR analysis of whole blood.
Strains of laboratory mice bearing a germline deletion of some T cell receptor V beta genes have proven useful in a variety of studies of T cell receptor function. Analysis of genetic crosses between deleted and wild type strains can provide information about the relevance of genes located within the deletion to specific T cell responses. Existing techniques for genotyping offspring of such crosses usually involve flow cytometric analysis which may not be available to all laboratories. Recent nucleotide sequence data indicate the presence of two restriction enzyme site polymorphisms in the closely linked V beta 1 gene which discriminate between deleted and wild-type strains. Amplification of a DNA segment containing the diagnostic sites by polymerase chain reaction followed by restriction enzyme digestion of the product offers a simple and rapid method for genotyping animals.
Spinella
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Veterans Administration Medical Center, Memphis, TN 38104.
D000818:Animals; D001483:Base Sequence; D001769:Blood; D004247:DNA; D004587:Electrophoresis, Agar Gel; D005821:Genetic Techniques; D005838:Genotype; D006239:Haplotypes / Q000235:genetics*; D051379:Mice; D008811:Mice, Inbred DBA; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D015183:Restriction Mapping
D011948:Receptors, Antigen, T-Cell; D004247:DNA