Rapid detection of DNA sequence variants by conformation-sensitive capillary electrophoresis

Genomics. 2000 Nov 15;70(1):34-40. doi: 10.1006/geno.2000.6354.

Abstract

The identification of novel sequence variants, which may be either disease-causing mutations or silent polymorphisms, in large numbers of samples is becoming the rate-limiting step in associating diseases with specific genes. This is particularly true in light of the imminent arrival of the complete reference sequence of the human genome. A number of techniques have been developed to analyze DNA samples for sequence variants rapidly. We describe a new technique, capillary-based conformation-sensitive gel electrophoresis (capillary CSGE) that transfers mutation detection from acrylamide gel to capillary electrophoresis. Capillary CSGE was able to detect 7/7 short insertion/deletions and 16/22 base substitutions in a series of random single-nucleotide polymorphisms and known variants in the lipoprotein lipase and BRCA2 genes. This technique has the potential to screen many megabases of DNA in a single day.

MeSH terms

  • BRCA2 Protein
  • DNA
  • Electrophoresis, Capillary / methods*
  • Genetic Variation*
  • Humans
  • Lipoprotein Lipase / genetics*
  • Neoplasm Proteins / genetics*
  • Nucleic Acid Conformation
  • Sequence Analysis, DNA
  • Transcription Factors / genetics*

Substances

  • BRCA2 Protein
  • Neoplasm Proteins
  • Transcription Factors
  • DNA
  • Lipoprotein Lipase