EXPONENTIAL BASE-3 NUCLEIC ACID AMPLIFICATION WITH REDUCED AMPLIFICATION TIME USING NESTED OVERLAPPING PRIMERS

摘要

Methods and primer set compositions that provide highly efficient nucleic acid amplification. 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. The method is based on the use of a set of nested primers an outer primer which comprises modified bases, e.g. 2-amino-adenine or 2-thio-thymine, and an inner primer which comprises a single-stranded target-binding region and a double stranded region comprising a flap sequence which is not complementary to target and a region which is complementary to the outer primer. The modified bases improve PCR cycle times, as compared to those observed in the absence of modified bases by allowing the outer primer to bind the target. The method and composition may also employ and contain, respectively, a third intermediate primer which has a structure as the inner primer.