Patent ID: 6534269
Filing Date: 2003-03-18
Classification: C12Q

Abstract:
A pyrophosphorolysis activated polymerization (PAP) method of synthesizing a desired nucleic acid strand on a nucleic acid template strand which comprises serially(a) annealing to the template strand a complementary activatable oligonucleotide P* that has a non-extendable 3â€²-deoxynucleotide at its 3â€² terminus and that has no nucleotides at or near its 3â€² terminus that mismatch the corresponding nucleotides on the template strand, so that the terminal 3â€²-deoxynucleotide is hybridized to the template strand when the oligonucleotide P* is annealed, (b) pyrophosphorolyzing the resulting duplex with pyrophosphate and an enzyme that has pyrophosphorolysis activity and activates the oligonucleotide P* by removal of the hybridized terminal 3â€²-deoxynucleotide, and (c) polymerizing by extending the activated oligonucleotide P* on the template strand in presence of four nucleoside triphosphates and a nucleic acid polymerase to synthesize the desired nucleic acid strand.