Analysis of Unstable Triplet Repeats Using Small-Pool Polymerase Chain Reaction
| Abstract |
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Small-pool polymerase chain reaction (PCR) constitutes the PCR amplification of a trinucleotide repeat in multiple small pools
of input DNA containing in the order of from 0.5 to 200 genome equivalents. Products are resolved by agarose gel electrophoresis
and detected by Southern blot hybridization under conditions that allow the identification of products derived from single-input
molecules. The method allows the detailed quantification of the degree of repeat-length variation in a given sample, including
the detection of common variants and those alleles present only in a small subset of cells. Detailed analysis of repeat dynamics
is essential for a complete understanding of the molecular mechanisms that generate diversity and lead to disease in the unstable
trinucleotide DNA repeat disorders.
Affiliation(s): (2) Division of Molecular Genetics, Institute of Biomedical and Life Sciences, Anderson College Complex, University of Glasgow, Glasgow, UK
(3) Departments of Biochemistry & Molecular Biology and Pediatrics, University of Oklahoma Health Sciences Center, Oklahoma City, OK
(4) Division of Molecular Genetics, Institute of Biomedical and Life Sciences, University of Glasgow, Anderson College Complex, Glasgow, UK
(3) Departments of Biochemistry & Molecular Biology and Pediatrics, University of Oklahoma Health Sciences Center, Oklahoma City, OK
(4) Division of Molecular Genetics, Institute of Biomedical and Life Sciences, University of Glasgow, Anderson College Complex, Glasgow, UK
Book Title: Trinucleotide Repeat Protocols
Series: Methods in Molecular Biology | Volume: 277 | Pub. Date: Jun-18-2004 | Page Range: 61-76 | DOI: 10.1385/1-59259-804-8:061
Subject: Cell Biology
Key Words: Mutation - polymerase chain reaction - trinucleotide repeat - myotonic dystrophy - Huntington’s disease - Friedreich ataxia - spinocerebellar ataxia - small-pool PCR - unstable DNA - expansion - repeat dynamics - Southern blot - triplet repeat - sperm - microsatellite
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