Mutation Typing Using Electrophoresis and Gel-Immobilized AcryditeTM Probes
Author(s) -
Mary Kenney,
Satyajit Ray,
T. Christian Boles
Publication year - 1998
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/98253pf01
Subject(s) - oligonucleotide , gel electrophoresis , microbiology and biotechnology , typing , nucleic acid , electrophoresis , polyacrylamide gel electrophoresis , nucleic acid thermodynamics , temperature gradient gel electrophoresis , biology , peptide nucleic acid , polymerase chain reaction , chemistry , dna , chromatography , gene , biochemistry , genetics , base sequence , enzyme , 16s ribosomal rna
A new electrophoresis technology for hybridization-based sequence detection and mutation typing is described. Intrinsic to this approach is copolymerization of specially modified oligonucleotide probes directly into polyacrylamide gels. Electrophoresis of single-stranded samples through gels containing specific immobilized probes results in hybridization-mediated capture of complementary targets. By increasing gel temperature or including denaturants in the buffer, the method can be used to type single-nucleotide polymorphisms. The method can easily be adapted to type mutations in PCR-amplified samples. Acrydite gel technology will also be useful for many other applications, including hybridization-based diagnostics, analysis of gene expression and purification of nucleic acids from biological samples.
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