title: Rolling circle amplification of DNA immobilized on solid surfaces and its application to multiplex mutation detection.
abstract: 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.
authors: Center for Advanced Biotechnology, Boston University, MA 02215, USA.
authors: A
authors: A
authors: Hatch
authors: T
authors: T
authors: Sano
authors: J
authors: J
authors: Misasi
authors: C L
authors: CL
authors: Smith
mesh_terms: D004247:DNA; D017931:DNA Primers; D009154:Mutation; D021141:Nucleic Acid Amplification Techniques; D015347:RNA Probes; D013698:Templates, Genetic
chemical_list: D017931:DNA Primers; D015347:RNA Probes; D004247:DNA