CPC C12Q 1/6848 (2013.01) [C12Q 1/6806 (2013.01); C12Q 1/6809 (2013.01); C12Q 1/6811 (2013.01); C12Q 1/6855 (2013.01); C12Q 1/686 (2013.01); C12Q 2525/101 (2013.01); C12Q 2525/186 (2013.01)] | 10 Claims |
1. A method of amplifying a target nucleic acid in a sample with reduced primer-primer interaction comprising:
(a) a primer extension step, wherein the sample is contacted with: (i) a nucleic acid polymerase, and (ii) a first primer which is a single-stranded oligonucleotide comprising at least one modified nucleotide that stalls nucleotide incorporation by the polymerase, the first primer comprising two arms complementary to a target sequence separated by a linker sequence non-complementary to the target sequence, such that a gap is created between the ends of the two arms when the first primer hybridizes to the target nucleic acid, wherein the polymerase fills the gap between the ends of the two arms; and
(b) an exponential amplification step wherein the sample is contacted with a second primer complementary to the primer extension product and a polymerase tolerant of the modified nucleotide.
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