Patent ID: 9637783
Date: 2017-05-02
CPC Classifications: C12N,C12P,C12Q,C12Y

Claim:
1. A process for increasing the number of copies of an oligonucleotide having an initial oligonucleotide sequence, wherein said oligonucleotide contains one or more non-standard nucleotides that carry a heterocycle selected from the group consisting of wherein R is the point of attachment of said heterocycle to the 2′-deoxyribose backbone in said chain, said process comprising: (a) dissolving said oligonucleotide in an aqueous mixture containing a thermostable DNA polymerase, nucleoside triphosphates that are Watson-Crick complementary to the nucleotides in said oligonucleotide, a first oligonucleotide primer that is sufficiently complementary to a segment at or near the 3′-end of said oligonucleotide that it hybridizes to said oligonucleotide, and a second oligonucleotide primer that has a sequence substantially identical to a portion of said oligonucleotide at or near its 5′-end, (b) incubating said mixture at a temperature where said polymerase extends the first oligonucleotide primer to give an extension product that is substantially complementary to said oligonucleotide, wherein said extension product forms a duplex with said oligonucleotide, such that said extension product, when it is separated from said oligonucleotide, can hybridize with the second primer, (c) then increasing the temperature of said mixture to a temperature sufficient to separate said oligonucleotide from said extension product, (d) then lowering the temperature of said mixture to a temperature at which the first oligonucleotide primer can hybridize to said oligonucleotide and the second oligonucleotide primer can hybridize to said extension product, and (e) repeating steps (b) through (d).