US 11,713,489 B2
Methods for temperature-mediated nested polymerase chain reaction
William M. Nelson, Rockville, MD (US); Kyle Armantrout, Los Angeles, CA (US); and Tracy Calvin Fecteau, Ellicott City, MD (US)
Assigned to Tetracore, Inc., Rockville, MD (US)
Filed by Tetracore, Inc., Rockville, MD (US)
Filed on Oct. 26, 2020, as Appl. No. 17/80,085.
Application 17/080,085 is a continuation of application No. 15/235,596, filed on Aug. 12, 2016, granted, now 10,837,065.
Claims priority of provisional application 62/205,248, filed on Aug. 14, 2015.
Prior Publication US 2021/0363568 A1, Nov. 25, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/689 (2018.01); C12Q 1/6844 (2018.01); C12Q 1/70 (2006.01)
CPC C12Q 1/689 (2013.01) [C12Q 1/6844 (2013.01); C12Q 1/701 (2013.01)] 18 Claims
OG exemplary drawing
 
1. A method for amplifying nucleic acid, comprising:
preparing a reaction mixture comprising at least one target nucleic acid, a first set of primers, and a second set of primers; and
processing the reaction mixture in a thermocycler, the processing comprising:
a first phase comprising heating the reaction mixture to a first temperature and cooling the reaction mixture to a second temperature repeatedly for a first plurality of cycles; and
a second phase comprising heating the reaction mixture to a third temperature and cooling the reaction mixture to a fourth temperature repeatedly for a second plurality of cycles, wherein the fourth temperature is gradually decreased over a predetermined number of cycles during the second phase;
wherein the first set of primers are configured to have melting temperatures around the second temperature and the second set of primers configured to have melting temperatures around the fourth temperature; and
wherein the melting temperatures of the first set of primers are higher than those of the second set of primers.