| 摘要 |
PURPOSE:To accurately determine the sequence of DNA bases, by carrying out the synthesis of a complementary DNA using as a primer a single chain DNA fragment loaded on a specific site of a double chain DNA contg. a specimen DNA and also using the specimen DNA as a template. CONSTITUTION:A double chain DNA contg. a specimen DNA and a single chain DNA fragment acting as a primer are allowed to coexist in the presence of recA protein or its analogous protein and ATP to load the single chain DNA fragment on a site adjacent to the specimen DNA of the double chain DNA in a form of D loop. Subsequently, the synthesis of a complementary DNA is performed on four kinds of ddNTPs the specimen DNA of the double chain DNA as a template in the presence of four kinds of dNTPs corresponding to the ddNTPs and 2', 3'-ddNTP corresponding to one kind of the base of the dNTPs. DNA molecules having different chain lengths are produced by this process. After released from the double chain, the formed DNA molecules are developed by a gel electrophoresis method and each base corresponding to each band is identified.
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