The present invention is directed to novel methods of synthesizing
multiple copies of a target nucleic acid sequence which are autocatalytic
(i.e., able to cycle automatically without the need to modify reaction
conditions such as temperature, pH, or ionic strength and using the
product of one cycle in the next one). In particular, the present
invention discloses a method of nucleic acid amplification which is
robust and efficient, while reducing the appearance of side-products. The
method uses only one primer, the "priming oligonucleotide," a promoter
oligonucleotide modified to prevent polymerase extension from its
3'-terminus and, optionally, a means for terminating a primer extension
reaction, to amplify RNA or DNA molecules in vitro, while reducing or
substantially eliminating the formation of side-products. The method of
the present invention minimizes or substantially eliminates the emergence
of side-products, thus providing a high level of specificity.
Furthermore, the appearance of side-products can complicate the analysis
of the amplification reaction by various molecular detection techniques.
The present invention minimizes or substantially eliminates this problem,
thus providing an enhanced level of sensitivity.