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METHODS OF DETECTING SEQUENCE DIFFERENCES

  • US 20110300537A1
  • Filed: 05/02/2011
  • Published: 12/08/2011
  • Est. Priority Date: 06/28/2002
  • Status: Abandoned Application
First Claim
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1. A method of determining, for a given nucleic acid sample, the identities of the nucleotides at a set of known polymorphic sites to be interrogated, said method comprising:

  • a) subjecting to an amplification regimen, a population of primer extension products generated from a nucleic acid sample, each primer extension product comprising a member of a set of tag sequences, which tag sequence specifically corresponds to the presence of one specific nucleotide at a known polymorphic site, wherein said amplification regimen is performed using one upstream amplification primer for each sequence comprising a known polymorphic site to be interrogated, and a set of distinguishably labeled downstream amplification primers, each member of said set of downstream amplification primers comprising a said tag sequence comprised by a member of said population of primer extension products and a distinguishable label that specifically corresponds to the presence of a specific nucleotide at said polymorphic site, and wherein said upstream amplification primers are selected such that each polymorphic site of said set of known polymorphic sites to be interrogated corresponds to a distinctly sized amplification product;

    b) detecting incorporation of a distinguishable label in distinctly sized amplification products, thereby to determine the identity of the nucleotide at each said polymorphic site.

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