Method for detecting DNA basic group mutation

Method for detecting DNA basic group mutation

  • CN 101,838,688 A
  • Filed: 04/07/2008
  • Published: 09/22/2010
  • Est. Priority Date: 04/07/2008
  • Status: Active Application
First Claim
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1. one kind is detected the method that whether has base mutation among the DNA, may further comprise the steps successively:

  • 1) preparation strand polynucleotide K1, the normal sequence of described K1 and DNA to be measured is complementary fully;

    2) preparation strand polynucleotide K2, the middle portion of described K2 has the DNAzyme activity, and K2 and K1 have at least 15nt complementary fully, and include the DNAzyme sequence of 10nt at least, and K2 and K1 at least respectively have 6nt not complementary end to end;

    3) preparation DNAzyme substrate S uses fluorescein-labelled S;

    The DNA part of substrate S is complementary fully with K2, and the restriction enzyme site that comprises two special RNA bases is arranged in the middle of the substrate S;

    4) K1 and K2 are mixed, add substrate S then, the compound that adds DNA to be measured and following formula (I), the control systems of the compound of dna fragmentation that adding is made up of the normal sequence of DNA to be measured and following formula (I) is set simultaneously, determines as follows whether DNA to be measured exists sudden change;

    if the I of DNA reaction system to be measured 424nm/ I 527nmSmaller or equal to 1.595 ±

    0.06 of control systems, there is base mutation in DNA to be measured;

    If the I of DNA reaction system to be measured 424nm/ I 527nmEqual 3.08 ±

    0.1 of control systems, there is not base mutation in DNA to be measured;

    Described I 424nmFor emission wavelength is the fluorescence intensity of the emission peak of 424nm;

    Described I 527nmFor emission wavelength is the fluorescence intensity of the emission peak of 527nm. Formula (I);

    In the formula (I), m=1~

    10, n=2~

    100;

    R 1, R 2, R 3, R 4=C XH 2X+1Or-O (C XH 2X+1), x=1~

    10;

    R 5=C XH 2X+1, x=1~

    3;

    R 6=Br, Cl, I, CF 3COO or CH 3COO.

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