title: An enhanced polymerase chain reaction assay to detect pre- and full mutation alleles of the fragile X mental retardation 1 gene.
abstract: Several diagnostic strategies have been applied to the detection of FMR1 gene repeat expansions in fragile X syndrome. Here, we report a novel polymerase chain reaction-based strategy using the Expand Long Template PCR System (Roche Diagnostics, Mannheim, Germany) and the osmolyte betaine. Repeat expansions up to approximately 330 CGGs in males and up to at least approximately 160 CGGs in carrier women could be easily visualized on ethidium bromide agarose gels. We also demonstrated that fluorescence analysis of polymerase chain reaction products was a reliable tool to verify the presence of premutation and full mutation alleles both in males and in females. This technique, primarily designed to detect premutation alleles, can be used as a routine first screen for expanded FMR1 alleles.
authors.affiliation: Dipartimento di Genetica Biologia e Biochimica, Università degli Studi di Torino, via Santena 19, 10126 Torino, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Saluto
authors.forename: Alessandro
authors.initials: A
authors.lastname: Brussino
authors.forename: Flora
authors.initials: F
authors.lastname: Tassone
authors.forename: Carlo
authors.initials: C
authors.lastname: Arduino
authors.forename: Claudia
authors.initials: C
authors.lastname: Cagnoli
authors.forename: Patrizia
authors.initials: P
authors.lastname: Pappi
authors.forename: Paul
authors.initials: P
authors.lastname: Hagerman
authors.forename: Nicola
authors.initials: N
authors.lastname: Migone
authors.forename: Alfredo
authors.initials: A
authors.lastname: Brusco
mesh_terms: D000483:Alleles; D001622:Betaine; D004252:DNA Mutational Analysis; D005260:Female; D005456:Fluorescent Dyes; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000235:genetics*; D006801:Humans; D008297:Male; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D019680:Trinucleotide Repeat Expansion
chemical_list: D001622:Betaine; D005456:Fluorescent Dyes; D051860:Fragile X Messenger Ribonucleoprotein 1