High-throughput screening for known mutations by automated analysis of single sequencing reactions

Biotechniques. 1998 May;24(5):832-5. doi: 10.2144/98245dt04.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aromatase / chemistry
  • Aromatase / genetics*
  • Genetic Testing / instrumentation
  • Genetic Testing / methods*
  • Genotype
  • Humans
  • Leukocytes / enzymology
  • Mutation*
  • Polymerase Chain Reaction
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Sequence Analysis, DNA / instrumentation
  • Sequence Analysis, DNA / methods*

Substances

  • Aromatase