Modified Primers for Nucleic Acid Amplification and Detection

摘要

A method of nucleic acid amplification involving using a first modified primer which provides protection to the amplification product from exonuclease degradation and a second primer. The method provides a double stranded nucleic acid, one strand of which is degraded by a double strand nucleic acid specific exonuclease to form a single stranded nucleic acid, which is protected from exonuclease degradation.

主权项

1. A nucleic acid amplification method comprising steps of:
a) performing nucleic acid amplification on a sample, using a first primer including at least one modified nucleotide (“modified primer”) and a second primer, wherein the amplification provides a double stranded nucleic acid comprising a first strand comprising the modified primer and a downstream amplified region; and a second strand; and b) incubating the double stranded nucleic acid of a) with a 5′ to 3′ double stranded nucleic acid specific exonuclease which hydrolyses the second strand but does not hydrolyse the amplified region of the first strand, to provide a single stranded nucleic acid comprising the amplified region of the first strand.