Compositions of toehold primer duplexes and methods of use

摘要

Provided herein are primers and primer systems having improved specificity and kinetics over existing primers, and methods of use thereof.

主权项

1. A partially double-stranded primer comprised of first and second nucleic acid strands arranged into
(1) one double-stranded target-non-specific region, (2) one double-stranded target-specific region, and (3) one single-stranded target-specific region contributed to by the first nucleic acid strand, wherein the double-stranded target-non-specific region has a standard free energy that is within 10% of the standard free energy for the single-stranded target-specific region bound to a perfectly complementary region of a target nucleic acid, wherein the sequence of the first nucleic acid strand that contributes to region (1) is not perfectly complementary to and does not bind to the target nucleic acid, and wherein the sequences of regions (2) and (3) are complementary to and bind to the target nucleic acid.