EXPONENTIAL BASE-GREATER-THAN-BASE 2 NUCLEIC ACID AMPLIFICATION USING HIGH- AND LOW-TM PRIMERS

Described herein are methods and compositions that provide highly efficient nucleic acid amplification. The method employs pairs of primers that differ significantly in Tm and a novel temperature/time course characterized by a temperature pulse during denaturation that enables a high-Tm primer (but not a low-Tm primer) to anneal and prime the synthesis of an additional nucleic acid strand beyond the two strands synthesized in a cycle of classical PCR. In some embodiments, this allows a 3-fold or greater increase of amplification product for each amplification cycle and therefore increased sensitivity and speed over conventional PCR.