US 11,866,775 B2
Methods for isothermal molecular amplification with nanoparticle-based reactions
Oleg Gang, Setauket, NY (US); and Suchetan Pal, New York, NY (US)
Assigned to Brookhaven Science Associates, LLC, Upton, NY (US)
Filed by Brookhaven Science Associates, LLC, Upton, NY (US)
Filed on Oct. 7, 2020, as Appl. No. 17/065,255.
Application 17/065,255 is a division of application No. 15/983,804, filed on May 18, 2018, abandoned.
Claims priority of provisional application 62/508,682, filed on May 19, 2017.
Prior Publication US 2021/0024990 A1, Jan. 28, 2021
Int. Cl. C12Q 1/6862 (2018.01); B82Y 5/00 (2011.01); C12Q 1/6853 (2018.01); C12Q 1/6844 (2018.01)
CPC C12Q 1/6862 (2013.01) [B82Y 5/00 (2013.01); C12Q 1/6844 (2013.01); C12Q 1/6853 (2013.01); C12Q 2545/101 (2013.01)] 18 Claims
 
1. A method for amplifying a molecular signal of an Input-Oligonucleotide, the method comprising
a) providing a plurality of Particle-A protected by oligonucleotide origami, wherein Particle-A is functionalized with a plurality of Oligonucleotide-1, wherein a sequence in Oligonucleotide 1 complementary to a sequence in a plurality of Oligonucleotide 2 hybridize to form Duplex 1-2, and sequence in Oligonucleotide 2 complementary to a sequence in oligonucleotide origami hybridize;
b) providing a plurality of Particle-B, wherein Particle-B is functionalized with a plurality of Oligonucleotide-3, wherein a sequence in Oligonucleotide-3 complementary to a sequence in a plurality of Stored-Input-Oligonucleotide hybridize to form Duplex 3-Y; and
c) mixing the Input-Oligonucleotide, plurality of origami protected Particle-A and plurality of Particle-B, in any order, wherein a cascade reaction occurs so that:
i) sequence in the Input-Oligonucleotide complementary to sequence in Oligonucleotide-2 hybridize to form Duplex Y-2, thereby removing Oligonucleotide-2 and the oligonucleotide origami, and exposing Oligonucleotide-1 on Particle A,
ii) sequence in Oligonucleotide-1 complementary to sequence in Oligonucleotide-3 hybridize to form Duplex 1-3, thereby releasing the Stored-Input-Oligonucleotide hybridized to the sequence in Oligonucleotide-3,
iii) sequence in the Stored-Input Oligonucleotide released in ii) complementary to sequence in Oligonucleotide-2 hybridize to form Duplex Y-2, thereby removing Oligonucleotide-2 and the oligonucleotide origami, and exposing Oligonucleotide-1 on Particle A to hybridize to complementary sequence in Oligonucleotide-3 to form Duplex 1-3, thereby releasing the Stored-Input-Oligonucleotide hybridized to the sequence in Oligonucleotide-3, and,
iv) where (iii) reoccurs until the cascade reaction terminates, when Particle A are fully deprotected.