| 摘要 |
<P>PROBLEM TO BE SOLVED: To provide a method for detecting single nucleotide variation, by solving such a problem that an apparatus for detecting the single nucleotide variation in a deoxyribonucleic acid by polyacrylamide gel electrophoresis becomes large in conventional methods, because of requiring temperature control. <P>SOLUTION: In this method, a single-stranded DNA into which deletion is introduced for the purpose of forming a single-stranded loop from a midway of a double-stranded DNA is added to a reaction liquid after completion of PCR reaction as a hybridization probe and subjected once to thermal melting, annealing, and polymerase extension reaction. A hybrid molecule which is hybridized with the single-stranded DNA having the deletion and in which the single-stranded loop is formed is sharply decreased in mobility when compared with a normal double-stranded DNA in the unmodified polyacrylamide gel electrophoresis. The single-stranded DNA hybridization probe used for forming the loop is designed so that a deletion part is adjacent to a position at which the single nucleotide variation is expected to occur, and therefore the single nucleotide variation is detected as a difference of the mobility due to a change of length of the loop by the polyacrylamide gel electrophoresis with a mini-gel at room temperature. <P>COPYRIGHT: (C)2007,JPO&INPIT |
