Patent ID: 6074818
Filing Date: 2000-06-13
Classification: C12Q,Y10S

Abstract:
A method for generating a profile characteristic of an unspecified nucleic acid sequence in a sample, which sequence is a template, which method comprises:a) treating the template with an oligonucleotide primer, wherein the primer has a double stranded stem of 2 complementary annealed base pairs, the double stranded stem extending to the 5' end of the primer and the other end of the stem extending as the single strand of at least 3 bases to the 3' end of the primer, and intermediate to the 5' and 3' ends of the stems a single stranded loop of 3 or 4 bases, the amount of primer being in excess over the amount of said nucleic acid, the amounts being in mass, and allowing said primer to anneal to multiple sites on each strand of said nucleic acid, each site being substantially complementary to the nucleotide of which said primer is constituted, thus, forming a multiplicity of a set of primed templates,b) treating said primed templates with a nucleic acid polymerase, thereby generating a multiplicity of extension strands, the extension strands comprising the primer in combination with a sequence of nucleotides that is substantially complementary to the templates, and extending along the template strands to either the 5' end of the template strands or the next primed site on the template, the number of extension strands thereby corresponding substantially to the number of primed sites,c) denaturing the extension strands from the template strands, thereby generating corresponding single-stranded extension strands and templates,d) allowing said primer to anneal to the extension strands and to the template strands,e) repeating steps (a), (b), (c) and (d), thereby generating nucleic acid fragments which are characteristic and unique for said nucleic acid of unspecified sequence,f) separating said fragments, andg) generating a characteristic profile of fragments from said sequence of nucleic acid in the sample.