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Compositions and methods for multiplex analysis of polynucleotides

  • US 20040229253A1
  • Filed: 02/18/2004
  • Published: 11/18/2004
  • Est. Priority Date: 02/18/2003
  • Status: Abandoned Application
First Claim
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1. A method of analyzing a polynucleotide sample for one or more target sequences, comprising the steps of:

  • contacting a polynucleotide sample suspected of comprising one or more target sequences with;

    (i) a first signal probe which is capable of hybridizing to at least a portion of a first target sequence and producing a first detectable signal when hybridized thereto;

    (ii) a first quencher probe which is capable of hybridizing in quenching proximity to the first signal probe and quenching the signal of the first signal probe when hybridized in quenching proximity thereto, said first quencher probe having a Tm below that of the first signal probe;

    (iii) at least a second signal probe which is capable of hybridizing to at least a portion of a second target sequence and producing a second detectable signal when hybridized thereto; and

    (iv) an optional second quencher probe which is capable of hybridizing in quenching proximity to the second signal probe and quenching the signal of the second signal probe when hybridized in quenching proximity thereto, said optional second quencher probe having a Tm below that of the second signal probe;

    monitoring the detectable signals of the signal probes as a function of temperature; and

    determining therefrom the presence or absence of one or more target sequences in said polynucleotide sample.

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