Methods for making nucleic acids

摘要

Nucleic acids are made by converting a primed single-stranded DNA to a double-stranded DNA by a method comprising the step contacting the single-stranded DNA with a DNA polymerase having 5' exonuclease activity under conditions whereby the DNA polymerase converts the-single stranded DNA to the double-stranded DNA, wherein the single-stranded DNA is primed with oligonucleotide primer comprising a sequence complementary to the 3' end of the single-stranded DNA, and at least one of the 5' end of the primer and the single-stranded DNA comprises an RNA polymerase promoter joined to an upstream (5') flanking moiety which protects the promoter from the 5' exonuclease activity of the DNA polymerase.