title: The facile detection of the nt 1226 mutation of glucocerebrosidase by 'mismatched' PCR.
abstract: The most common Gaucher disease-producing mutation among Ashkenazi Jews is an A----G substitution at cDNA nt 1226 (genomic nt 5841). We describe a simple method for detecting this mutation both in genomic DNA and in cDNA by performing polymerase chain reaction (PCR) using a 5'-primer mismatched at one nucleotide so as to create an Xho I restriction site. When the mutation is present. the 105 bp fragment formed is cleaved to 89 and 16 nt fragments. The 89 bp fragment is easily visualized on a gel making it possible to distinguish individuals who do not have the mutation from heterozygotes and homozygotes.
authors.affiliation: Scripps Clinic and Research Foundation, Department of Molecular and Experimental Medicine, La Jolla, CA 92037.
authors.forename: E
authors.initials: E
authors.lastname: Beutler
authors.forename: T
authors.initials: T
authors.lastname: Gelbart
authors.forename: C
authors.initials: C
authors.lastname: West
mesh_terms: D001483:Base Sequence; D015252:Deoxyribonucleases, Type II Site-Specific; D005776:Gaucher Disease / Q000201:enzymology*; D005962:Glucosylceramidase / Q000235:genetics*; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction
chemical_list: C034510:CTCGAG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; D005962:Glucosylceramidase