US 12,077,811 B2
Compositions of toehold primer duplexes and methods of use
David Yu Zhang, Cambridge, MA (US); and Peng Yin, Cambridge, MA (US)
Assigned to President and Fellows of Harvard College, Cambridge, MA (US)
Filed by President and Fellows of Harvard College, Cambridge, MA (US)
Filed on Feb. 5, 2021, as Appl. No. 17/169,145.
Application 14/553,165 is a division of application No. 13/882,231, granted, now 9,284,602, previously published as PCT/US2011/058178, filed on Oct. 27, 2011.
Application 17/169,145 is a continuation of application No. 16/017,570, filed on Jun. 25, 2018, abandoned.
Application 16/017,570 is a continuation of application No. 14/553,165, filed on Nov. 25, 2014, granted, now 10,036,059.
Claims priority of provisional application 61/407,291, filed on Oct. 27, 2010.
Prior Publication US 2021/0388430 A1, Dec. 16, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/68 (2018.01); C07H 21/02 (2006.01); C12Q 1/6832 (2018.01); C12Q 1/6848 (2018.01); C12Q 1/6853 (2018.01); C12Q 1/686 (2018.01)
CPC C12Q 1/6853 (2013.01) [C07H 21/02 (2013.01); C12Q 1/6832 (2013.01); C12Q 1/6848 (2013.01); C12Q 1/686 (2013.01)] 15 Claims
 
1. A system comprising a target nucleic acid and a nucleic acid comprising:
(1) a first double-stranded region comprising a sequence that is complementary to a first portion of the target nucleic acid,
(2) a second double-stranded region that is substantially non-complementary to the target nucleic acid, and
(3) a single-stranded region comprising a sequence that is complementary to a second portion of the target nucleic acid;
wherein the second double-stranded region has a concentration-adjusted standard free energy that is within 10% of the concentration-adjusted standard free energy for the single-stranded region bound to the second portion of the target nucleic acid.