AMPLIFICATION METHOD FOR POLYNUCLEOTIDE ASSAYS AMPLIFICATION METHOD FOR POLYNUCLEOTIDE ASSAYS

摘要

A method is disclosed for producing multiple copies of a primary polynucleotide sequence located at the 3 min terminus of a polynucleotide. The method comprises (a) forming in the presence of nucleoside triphosphates and template-dependent polynucleotide polymerase an extension of a primary polynucleotide sequence hybridized with a template sequence of a single stranded pattern polynucleotide comprising two or more template sequences each containing one or more cleavable sites (b) cleaving into fragments said extension at cleavable sites in the presence of means for specifically cleaving said cleavable sites when said extension is hybridized with said template sequence, (c) dissociating said fragments, (d) hybridizing said fragments with single stranded pattern polynucleotide, and repeating steps (a)-(d). Steps (a)-(d) may be conducted simultaneously or wholly or partially sequentially. The method may be applied in the detection of a polynucleotide analyte in a sample suspected of containing such analyte to facilitate such detection. Also disclosed are compositions for conducting the method of the invention.