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

Currently, there is a need for practical, accurate and cost-efficient tests to comprehensively scan human genes for disease-related DNA sequence varia tion. 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 co mprises a size separation step followed by denaturing gradient gel electrop horesis (DGGE), and allows single base pair changes to be distinguished amo ng multiple DNA fragments in parallel. Here, we describe the rapid design o f TDGS tests and its application to mutation identification in several larg e human cancer genes. (C) 1999 Elsevier Science B.V. All rights reserved.