Plant & Animal Genomes XIII Conference

January 15-19, 2005
Town & Country Convention Center
San Diego, CA

Isothermal Amplified From The Limited Quantities Of DNA Samples Using Novel Random Primers.

Hirokazu Takahashi¹ , Seigo Kuwasaki¹ , Yoshiataka Suetsugu¹ , Junko Narukawa¹ , Kazuei Mita¹ , Shigeru Sugiyama² , Toshio Ohtani² , Kimiko Yamamoto¹

¹  National Institute of Agrobiological Sciences, 1-2 Owashi, Tsukuba, Ibaraki 305-8634, Japan

²  National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan

Frequently, molecular analyses require certain amounts of DNA, although there are many cases that only limited amount of DNA samples are available. When amplification reaction is performed on a whole genome with small amount of template DNA, amplification of non-specific DNA molecules besides target DNA is serious problem. In amplification reaction by multi-strand displacement amplification (MDA) method at relatively low temperature using random primer, non-specific amplicon derived from the direct DNA synthesis by primer-primer pairs are frequently generated. To eliminate the non-specific amplification problem, an improvement of the MDA method by using novel random primer was performed. The non-specific amplicon were not synthesized in negative control such as distilled water, because DNA polymerases could not use the novel random primer as a template of DNA synthesis. As well as the method using normal random primer, amplified DNA was suitable to use in many application of whole genome analysis. This method will be a useful tool in molecular analyses of DNA in the case of only limited amount of DNA is available.
This research is supported by fund of Promotion of Basic Research Activities for Innovative Bioscience from BRAIN.