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Two‐dimensional gene scanning: Exploring human genetic variability
Author(s) -
Vijg Jan,
van Orsouw Nathalie J.
Publication year - 1999
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(19990101)20:6<1239::aid-elps1239>3.0.co;2-t
Subject(s) - multiplex , biology , genetics , computational biology , polymerase chain reaction , gene , dna sequencing , multiplex polymerase chain reaction , population , temperature gradient gel electrophoresis , demography , sociology , 16s ribosomal rna
Current methods for mutation detection are not optimized for the generation of highly accurate data on multiple genes of hundreds of individuals in population‐based studies. Two‐dimensional gene scanning (TDGS) is a high‐resolution system for detecting mutational variants in multiple genes in parallel. TDGS is based on a combination of extensive multiplex polymerase chain reaction (PCR) and two‐dimensional (2‐D) DNA electrophoresis. The latter involves a size separation step followed by denaturing gradient gel electrophoresis (DGGE). TDGS tests for a number of large human disease genes have been designed, using a computer program to optimally position PCR primers around the relevant target sequences ( e.g. , exons) and evaluated using panels of samples with previously detected mutations. The results indicate a high sensitivity and specificity, equal to nucleotide sequencing, which is generally considered as the gold standard. Here, we describe the different components of the TDGS process and its potential application as a high‐throughput system for the systematic identification of human gene variants.