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 3'blocked
promoter oligonucleotide and optionally, a means for terminating a primer
extension reaction, to amplify RNA or DNA molecules in vitro, while
reducing or eliminating the formation of side products. The method of the
present invention minimizes or 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 eliminates this problem, thus providing an enhanced level of
sensitivity.