The present invention provides a nucleotide mutation detection method wherein primers having a nucleotide sequence complementary to part of the nucleic acids to be detected and also having a nucleotide sequence, which is complementary to the nucleotide sequence just upstream from a nucleotide site corresponding to the mutation site of the nucleotide sequence formed at the 3' end, added to the 5' end, are produced so that so that the nucleotide site corresponding to the mutation site of the nucleic acids to be detected including the nucleotide mutation is located within the nucleotide sequence formed at the 3' end after elongation; a target is produced by subjecting the primers to an elongation reaction using polymerase or Klenow enzyme; the target is then denatured to a single strand, and is subjected to a hybridization reaction with a probe that has the nucleotide complementary to the mutation site of the nucleic acids present in the 3' end region and then to a ligation reaction, whereby it is enable to determine and assay a nucleotide located at a specific position in a nucleotide sequence of DNA or RNA and to analyze rapidly a variety of mutations including single nucleotide mutations, short nucleotide tandem repeat mutations, nucleotide deletion mutations, nucleotide insertion mutations, translocation mutations and so on.

 
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