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Method for profiling nucleic acids of unknown sequence using arbitrary oligonucleotide primers

  • US 5,413,909 A
  • Filed: 01/19/1993
  • Issued: 05/09/1995
  • Est. Priority Date: 08/24/1990
  • Status: Expired due to Fees
First Claim
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1. A method for generating a pattern characteristic of at least one nucleic acid of unspecified sequence in a sample, which method comprises:

  • a) treating said nucleic acid with at least one oligonucleotide primer having an arbitrary sequence and of 5 to 25 nucleotides in length, 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 nucleotides 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) until there are generated nucleic acid fragments which are characteristic and unique for said nucleic acid of unspecified sequence,f) separating said fragments, andg) determining the characteristic pattern of fragments generated from said sequence of nucleic acid in the sample.

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