Mutation typing using electrophoresis and gel-immobilized Acrydite probes.
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.
Kenney
M
M
Mosaic Technologies, Inc., Boston, MA, USA.
Ray
S
S
Boles
T C
TC
D004252:DNA Mutational Analysis; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics; D004277:DNA, Single-Stranded / Q000032:analysis* / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005814:Genes, Viral / Q000235:genetics; D006678:HIV / Q000201:enzymology / Q000235:genetics; D006801:Humans; D009154:Mutation / Q000235:genetics; D009693:Nucleic Acid Hybridization; D009943:Organophosphorus Compounds / Q000737:chemistry*; D011110:Polymorphism, Genetic; D012194:RNA-Directed DNA Polymerase / Q000235:genetics; D012680:Sensitivity and Specificity
D015342:DNA Probes; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D009943:Organophosphorus Compounds; D012194:RNA-Directed DNA Polymerase