title: High-throughput screening for known mutations by automated analysis of single sequencing reactions.
abstract: This work shows that single sequencing reactions analyzed on an automated DNA sequencer can be an efficient way of screening PCR products for known mutations. We have analyzed a mutation in exon 10 of the human aromatase gene and show that an unambiguous genotype could be elucidated in more than 90% of the analyzed samples. Compared to analysis by full sequencing, 4 times more samples can be analyzed per gel, so that the sample capacity of the gel is approaching that of alternative gel-based methods for genotype analysis. Unlike many of these, the method offers direct identification of the variant sequence position and on-line analysis without the need of post-electrophoretic processing of the gel.
authors.affiliation: Institute of Cancer Research, University of Oslo, Norway.
authors.forename: T
authors.initials: T
authors.lastname: Kristensen
authors.forename: V N
authors.initials: VN
authors.lastname: Kristensen
authors.forename: A L
authors.initials: AL
authors.lastname: Børresen-Dale
mesh_terms: D001141:Aromatase / Q000737:chemistry / Q000235:genetics*; D005820:Genetic Testing / Q000295:instrumentation / Q000379:methods*; D005838:Genotype; D006801:Humans; D007962:Leukocytes / Q000201:enzymology; D009154:Mutation; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000295:instrumentation / Q000379:methods*
chemical_list: D001141:Aromatase