Novel methods of synthesizing multiple copies of a target nucleic acid
sequence which are autocatalytic are disclosed (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, methods of nucleic acid amplification are
disclosed which are robust and efficient, while reducing the appearance
of side-products. In general, the methods use priming oligonucleotides
that target only one sense of a target nucleic acid, 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-pro ducts. The disclosed
methods minimizes or substantially eliminate 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
disclosed methods minimize or substantially eliminate this problem, thus
providing enhanced levels of sensitivity.