Patent ID: 8871469
Filing Date: 2014-10-28
Classification: C07H,C12Q

Abstract:
1. A process for amplifying one or more oligonucleotides by enzymatic template-directed primed polymerization, wherein said process comprises (a) contacting in aqueous solution one or more pairs of primers with said oligonucleotide, a polymerase, and standard nucleoside triphosphates, and (b) incubating the mixture for a preselected length of time, wherein each of said primers has the formula: wherein X is selected from the group consisting of OH, O-phosphate, O-oligonucleotide, —NH 2 , and a phosphate or an amino group linked to a biotin or fluorescent tag, B is independently selected from the group consisting of adenine, thymine, guanine, cytosine, diaminopurine, uracil, A*, T*, G*, and C*, D is independently selected from the group consisting of A*, T*, G*, and C*, E is independently selected from the group consisting of A, T, G, and C, and K is independently selected from the group consisting of A, T, G, C, A*, T*, G*, and C*, wherein n is an integer from 4 to 25 and m is an integer from 2 to 10, wherein the first member of said pair is complementary to said oligonucleotide at a portion of it, and the second member of said pair is formally the same as a segment of the same oligonucleotide at a position between 10 and 1000 nucleotides in the 5′-direction from the portion, where A* does not contribute to the stability of a duplex when paired with T* but does when it is paired with thymine, T* does not contribute to the stability of a duplex when paired with A* but does when it is paired with adenine, G* does not contribute to the stability of a duplex when paired with C* but does when it is paired with cytosine, and C* does not contribute to the stability of a duplex when paired with G* but does when it is paired with guanine.