Amplification method for polynucleotide assays.

A METHOD FOR PRODUCING MULTIPLE COPIES OF A PRIMARY POLYUCLEOTIDE SEQUENCE LOCATED AT THE 3 '' TERM OF A POLYUCLEOTIDE IS DESCRIBED. THE METHOD CONSISTS OF (A) FORMING, IN THE PRESENCE OF NUCLEOSID THYPHOSPHATES AND POLYMERASE OF TEMPORARY DEPENDENT POLYUCLEOTIDE, AN EXTENSION OF A SEQUENCE OF PRIMARY NUCLEOTIDE HYBRIDIZED WITH A SEQUENCE OF TEMPLATE OF A NUCLEOTIDE PATRON, MADE OF A SIMPLE FLEECE, CONTAINING 2 OR MORE TEMPLATE SEQUENCES, WHICH CONTAIN EACH ONE OR MORE BREAKABLE SITES; (B) BREAKING INTO FRAGMENTS SUCH EXTENSION IN ROMPABLE SITES, IN THE PRESENCE OF MEANS TO SPECIFICALLY BREAK THOSE SUCH ROMPABLE SITES, WHEN SUCH EXTENSION IS HYBRIDIZED WITH SUCH TEMPLATE SEQUENCE; (C) SEPARATE SUCH FRAGMENTS; (D) HYBRIDIZE SUCH FRAGMENTS WITH PATTERN POLYUCLEOTIDE, MADE SINGLE FLEECE, AND REPEAT THE STAGES (A) - (D). STAGES (A) - (D) CAN BE PERFORMED SIMULTANEOUSLY OR SEQUENTIAL, PARTIALLY OR COMPLETELY. THE METHOD CAN BE APPLIED FOR THE LADETECTION OF A POLYUCLEOTIDE ANALYZ IN A SAMPLE, WHICH IS SUSPECTED CONTAINS SUCH ANALYZE TO FACILITATE SUCH DETECTION. COMPOSITIONS TO CARRY OUT THE METHOD OF THE INVENTION ARE ALSO DESCRIBED.