Nucleic acid amplification with allele-specific suppression of sequence variants

Nucleic acid amplification with allele-specific suppression of sequence variants

  • CN 102,272,333 A
  • Filed: 01/07/2010
  • Published: 12/07/2011
  • Est. Priority Date: 01/07/2009
  • Status: Active Application
First Claim
Patent Images

1. the method that needs variant of a selective amplification target sequence, described target sequence exists with multiple variant form, said method comprising the steps of:

  • A) in reaction mixture, provide the sample of at least a variant that may comprise described target sequence;

    B) provide can with first oligonucleotide of the multiple variant hybridization of described target sequence;

    C) provide can with second oligonucleotide of the multiple variant hybridization of described target sequence, wherein said second oligonucleotide at least partially in the base that comprises modification near 3 '"'"' terminal or one or more Nucleotide it;

    D) provide the 3rd oligonucleotide, the avidity that needs variant hybridization of itself and described target sequence is lower than the avidity that does not need variant hybridization with described target sequence, and is designed to the chain identical with described second oligonucleotide hybridization and is positioned at 0-60 the Nucleotide in the described second oligonucleotide downstream;

    E) provide the nucleic acid polymerase that does not have 5 '"'"'-3 '"'"' nuclease basically;

    F) make described reaction mixture carry out the polymerase chain reaction,Wherein do not need variant when hybridization when described the 3rd oligonucleotide and described target sequence, described the 3rd oligonucleotide significantly suppresses the extension of described nucleic acid polymerase to described second oligonucleotide, but when described the 3rd oligonucleotide and described target sequence need variant when hybridization, it does not suppress the extension of described nucleic acid polymerase to described second oligonucleotide basically.

View all claims
    ×
    ×

    Thank you for your feedback

    ×
    ×