Design and application of 2-D DGGE-based gene mutational scanning tests

Genet Anal. 1999 Feb;14(5-6):205-13. doi: 10.1016/s1050-3862(98)00028-x.

Abstract

Currently, there is a need for practical, accurate and cost-efficient tests to comprehensively scan human genes for disease-related DNA sequence variation. Two-dimensional gene scanning (TDGS) is a parallel mutation detection system, based on a combination of extensive multiplex PCR amplification ('PCR megaplex') and two-dimensional (2-D) DNA electrophoresis. The latter comprises a size separation step followed by denaturing gradient gel electrophoresis (DGGE), and allows single base pair changes to be distinguished among multiple DNA fragments in parallel. Here, we describe the rapid design of TDGS tests and its application to mutation identification in several large human cancer genes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • BRCA1 Protein / genetics
  • Carrier Proteins
  • Computer Simulation
  • DNA Mutational Analysis / methods*
  • Electrophoresis, Gel, Two-Dimensional
  • Genes, p53 / genetics
  • Humans
  • Models, Biological
  • MutL Protein Homolog 1
  • Neoplasm Proteins / genetics
  • Nuclear Proteins
  • Polymerase Chain Reaction / methods*
  • Retinoblastoma Protein / genetics
  • Temperature

Substances

  • Adaptor Proteins, Signal Transducing
  • BRCA1 Protein
  • Carrier Proteins
  • MLH1 protein, human
  • Neoplasm Proteins
  • Nuclear Proteins
  • Retinoblastoma Protein
  • MutL Protein Homolog 1