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filingDate 2013-04-16-04:00^^<http://www.w3.org/2001/XMLSchema#date>
inventor http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_98e7efb3c16941aace1328574fa29619
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publicationDate 2019-03-26-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber CN-109517888-A
titleOfInvention Nucleic acid amplification method using allele-specific reactive primers
abstract The present invention relates to a method for amplifying nucleic acid using allele-specific reactive primers (ASRP) designed to solve the problems of conventional allele-specific PCR, and more particularly, the present invention relates to a method for detecting target nucleic acid , comprising a DNA polymerase with proofreading activity and a base sequence complementary to a target nucleic acid, wherein the target nucleic acid is amplified in the presence of an ASRP modified so as to be located from a nucleotide that is not complementary to the 5' direction of the primer One or more modified nucleotides in the region of immediately adjacent nucleotides to the nucleotides at the 5' end of the primer that, when present at the 3' end, are uncomplemented by the DNA The proofreading activity of the polymerase is removed so that the nucleotides in the allele-specific reactive primers cannot be used as primers for the polymerase reaction. The detection method using ASRP according to the present invention is a technique with very high specificity due to the properties of ASRP and proofreading DNA polymerase, and can efficiently detect mutations (dots) including single nucleotide polymorphisms (SNPs). mutation, insertion, deletion). In addition, this method can also be used to detect the presence of CpG methylation after bisulfite treatment, or to amplify and detect target DNA starting from a desired nucleotide sequence in a DNA library.
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