Method for relative quantification of nucleic acid sequence, expression, or copy changes, using combined nuclease, ligation, and polymerase reactions

Method for relative quantification of nucleic acid sequence, expression, or copy changes, using combined nuclease, ligation, and polymerase reactions

  • CN 104,204,228 A
  • Filed: 02/14/2013
  • Published: 12/10/2014
  • Est. Priority Date: 02/14/2012
  • Status: Active Application
First Claim
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1. for the identification of the method that in sample, one or more target nucleotide sequences exist, it comprises:

  • The sample that may contain described one or more target nucleotide sequences is provided;

    One or more oligonucleotide probe groups are provided, every group comprises (a) and has the first oligonucleotide probe of target-specific part, (b) there is the second oligonucleotide probe of target-specific part, wherein described first and second oligonucleotide probes of probe groups are configured to hybridization adjacent one another are in described target nucleotide sequences, between described the first and second oligonucleotide probes, form and engage, and wherein, in probe groups, the described target-specific part of described the second oligonucleotide probe has the identical Nucleotide overlapping with described the first oligonucleotide probe at described joint,Hybridize under effective condition with its corresponding target nucleotide sequences of base specific mode (if being present in sample) on consecutive position at the first and second oligonucleotide probes that make probe groups, described sample is contacted with described one or more oligonucleotide probe groups, wherein, after hybridization, form flank at the described overlapping identical Nucleotide of the second oligonucleotide probe described in the joint that comprises described overlapping identical Nucleotide;

    Cut the described overlapping identical Nucleotide of described the second oligonucleotide probe with the enzyme with 5 '"'"' nuclease, thereby be released in the phosphoric acid of 5 '"'"' end of described the second oligonucleotide probe;

    The first and second oligonucleotide probes of described one or more oligonucleotide probe groups are joined together to form to the product sequence of connection at described joint;

    Detect the described connection product sequence in described sample;

    WithIdentify the existence of one or more target nucleotide sequences in described sample based on described detection.

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