A novel SNaPshot assay to detect the mdx mutation.
The mdx mouse is an animal model for Duchenne muscular dystrophy (DMD). In order to evaluate possible treatments and to carry out genetic studies, it is essential to distinguish between mice that carry the dystrophic (mutant) or wild-type (wt) allele(s). The current amplification-resistant mutation system (ARMS) assay is labor intensive and yields false negatives, which reduces its efficiency as a screening tool. An alternate assay based on single-nucleotide polymorphism (SNP) primer extension technology (i.e., SNaPshot) is described. The SNaPshot assay has been optimized to identify both wild-type and mutant alleles, providing a robust, potentially automatable assay for high-throughput analysis.
Budowle
Sarah A
SA
Department of Human Nutrition, Food and Exercise, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061, USA. sbudowle@vt.edu
Gonzalez
Suzanne
S
Budowle
Bruce
B
Eisenberg
Arthur J
AJ
Grange
Robert W
RW
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D004252:DNA Mutational Analysis; D017931:DNA Primers; D004259:DNA-Directed DNA Polymerase / Q000096:biosynthesis / Q000235:genetics; D054306:Dideoxynucleotides; D019075:Electrophoresis, Capillary; D005456:Fluorescent Dyes; D005784:Gene Amplification; D051379:Mice; D008810:Mice, Inbred C57BL; D018101:Mice, Inbred mdx; D008969:Molecular Sequence Data; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics*; D009154:Mutation; D020641:Polymorphism, Single Nucleotide; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000379:methods*
D017931:DNA Primers; D054306:Dideoxynucleotides; D005456:Fluorescent Dyes; D004247:DNA; D004259:DNA-Directed DNA Polymerase