Rapid design of denaturing gradient-based two-dimensional electrophoretic gene mutational scanning tests

Nathalie J. Van Orsouw, Rahul K. Dhanda, R. David Rines, Wendy M. Smith, Iakovos Sigalas, Charis Eng, Jan Vijg

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

With the current rapid pace at which human disease genes are identified there is a need for practical, cost-efficient genetic screening tests. Two-dimensional electrophoretic separation of PCR-amplified gene fragments on the basis of size and base pair sequence, in non-denaturing and denaturing gradient polyacrylamide gels respectively, provides a rapid parallel approach to gene mutational scanning. Accuracy of the denaturing gradient gel electrophoresis (DGGE) component of this system strongly depends on the design of the PCR primers and the melting characteristics of the fragments they encompass. We have developed a fully automated generally applicable procedure to generate optimal two-dimensional test designs at a minimum amount of time and effort. Designs were generated for the RB1, TP53, MLH1 and BRCA1 genes that can be readily implemented in research and clinical laboratories as low cost genetic screening tests.

Original languageEnglish (US)
Pages (from-to)2398-2406
Number of pages9
JournalNucleic Acids Research
Volume26
Issue number10
DOIs
StatePublished - May 15 1998
Externally publishedYes

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Genetic Testing
BRCA1 Gene
Genes
Costs and Cost Analysis
Polymerase Chain Reaction
Denaturing Gradient Gel Electrophoresis
p53 Genes
Base Pairing
Freezing
Research
polyacrylamide gels

ASJC Scopus subject areas

  • Genetics

Cite this

Rapid design of denaturing gradient-based two-dimensional electrophoretic gene mutational scanning tests. / Van Orsouw, Nathalie J.; Dhanda, Rahul K.; Rines, R. David; Smith, Wendy M.; Sigalas, Iakovos; Eng, Charis; Vijg, Jan.

In: Nucleic Acids Research, Vol. 26, No. 10, 15.05.1998, p. 2398-2406.

Research output: Contribution to journalArticle

Van Orsouw, Nathalie J. ; Dhanda, Rahul K. ; Rines, R. David ; Smith, Wendy M. ; Sigalas, Iakovos ; Eng, Charis ; Vijg, Jan. / Rapid design of denaturing gradient-based two-dimensional electrophoretic gene mutational scanning tests. In: Nucleic Acids Research. 1998 ; Vol. 26, No. 10. pp. 2398-2406.
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