Rapid Design of Denaturing Gradient-based Two-dimensional Electrophoretic Gene Mutational Scanning Tests

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1998 Jun 10

PMID 9580692

Citations 4

Authors

N J van Orsouw

R K Dhanda

R D Rines

W M Smith

I Sigalas

C Eng

J Vijg

Affiliations

Soon will be listed here.

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.

Citing Articles

Detection by denaturing gradient gel electrophoresis of pncA mutations associated with pyrazinamide resistance in Mycobacterium tuberculosis isolates from the United States-Mexico border region.

McCammon M, Gillette J, Thomas D, Ramaswamy S, Rosas I, Graviss E Antimicrob Agents Chemother. 2005; 49(6):2210-7.

PMID: 15917514 PMC: 1140546. DOI: 10.1128/AAC.49.6.2210-2217.2005.

Detection of rpoB mutations associated with rifampin resistance in Mycobacterium tuberculosis using denaturing gradient gel electrophoresis.

McCammon M, Gillette J, Thomas D, Ramaswamy S, Graviss E, Kreiswirth B Antimicrob Agents Chemother. 2005; 49(6):2200-9.

PMID: 15917513 PMC: 1140537. DOI: 10.1128/AAC.49.6.2200-2209.2005.

A highly accurate, low cost test for BRCA1 mutations.

van Orsouw N, Dhanda R, Elhaji Y, Narod S, Li F, Eng C J Med Genet. 1999; 36(10):747-53.

PMID: 10528853 PMC: 1734249. DOI: 10.1136/jmg.36.10.747.

Automated mutation analysis.

Ravine D J Inherit Metab Dis. 1999; 22(4):503-18.

PMID: 10407782 DOI: 10.1023/a:1005508324048.

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