Method For Obtaining Subtraction Polynucleotide

  • Published: Feb 26, 2009
  • Earliest Priority: Mar 01 2005
  • Family: 7
  • Cited Works: 0
  • Cited by: 0
  • Cites: 4
  • Sequences: 7
  • Additional Info: Full text Sequence
Abstract

The present invention provides a method for obtaining or amplifying a polynucleotide (a tester-specific polynucleotide), in which an amount existing in a sample (tester) is larger than the amount existing in another sample (driver), easily and within a short time as well as with high efficiency, a polynucleotide obtained (amplified) by such method, a method for identifying gene mutation in the tester, and a kit to be used in such methods. The present invention discloses a method for obtaining a tester-specific polynucleotide comprising the following steps: (1) forming a double-stranded polynucleotide consisting of a single-stranded polynucleotide derived from the tester and a single-stranded polynucleotide derived from the driver, which is a complementary strand with the single-stranded polynucleotide derived from the tester, by (i) performing hybridization with the single-stranded polynucleotide derived from the tester and the single-stranded polynucleotide derived from the driver, which can be the complementary strand with the single-stranded polynucleotide derived from the tester, or (ii) performing hybridization after mixing the single-stranded polynucleotide derived from the tester and the double-stranded polynucleotide derived from the driver and subjecting to thermal denaturation, and (2) removing the hybridized double-stranded polynucleotide by enzymatic treatment, and a method for amplifying a tester-specific polynucleotide further comprising a step: (3) amplifying a non-hybridized single-stranded polynucleotide derived from the tester. According to the present invention, a tester-specific polynucleotide can be obtained or amplified easily and within a short time as well as with high efficiency, and as a result, gene mutation in the tester can be identified with a high degree of accuracy.


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Document History
  • Publication: Feb 26, 2009
  • Application: Feb 27, 2006
    US US 81665206 A
  • Priority: Feb 27, 2006
    JP 2006303585 W
  • Priority: Mar 1, 2005
    JP 2005055279 A

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