Methods of preventing non-specific reactions of nucleotide sequences

摘要

Disclosed herein are methods of nucleic acid amplification, including methods of preventing non-specific reaction of a nucleotide sequence with a DNA modifying enzyme.

主权项

1. A method of preventing non-specific reaction of a nucleotide sequence with a DNA modifying enzyme, comprising:
designing an oligonucleotide that comprises from about 5 to about 50 nucleotides, wherein about 40% to 100% of the nucleotides comprise a sulfur atom and wherein the oligonucleotide is not a primer, and has a 3′ cap to prevent extension by a polymerase; and contacting the oligonucleotide with at least one nucleic acid modifying enzyme, wherein the nucleic acid modifying enzyme is a polymerase; and performing amplification in the presence of the oligonucleotide, a primer, and the polymerase, wherein the oligonucleotide reduces enzymatic activity of the polymerase at or below room temperature, but that reduces enzymatic activity to a lesser degree at the reaction temperature, which is above room temperature; and wherein the oligonucleotide reduces nonspecific products produced by the polymerase; further wherein said oligonucleotide is comprised of polyA bases.