Rolling circle amplification of DNA immobilized on solid surfaces and its application to multiplex mutation detection.
A new method of amplifying short DNA molecules immobilized on a solid support has been developed. This method uses a solid-phase rolling circle replication reaction, termed rolling circle amplification (RCA). The probe consists of a single-stranded DNA primer anchored at the 5' terminus to a solid support and a single stranded DNA template hybridized to the immobilized primer. Here, DNA ligase was used to circularize the template, and DNA polymerase I was used to extend the immobilized primer in a rolling circle replication reaction. This method was used to identify a known polymorphism in BRCA1 exon 5. These results demonstrate that RCA offers considerable promise to facilitate effective mutation screening of DNA using a solid-phase format.
Hatch
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Center for Advanced Biotechnology, Boston University, MA 02215, USA.
Sano
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Misasi
J
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Smith
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CL
D004247:DNA; D017931:DNA Primers; D009154:Mutation; D021141:Nucleic Acid Amplification Techniques; D015347:RNA Probes; D013698:Templates, Genetic
D017931:DNA Primers; D015347:RNA Probes; D004247:DNA