US 12,385,103 B1
Surface plasmon resonance genotyping for plant breeding
Ryan Manuszak, Johnston, IA (US); and Micheal Tuntland, Indianapolis, IN (US)
Assigned to PIONEER HI-BRED INTERNATIONAL, INC., , IA (US)
Filed by PIONEER HI-BRED INTERNATIONAL, INC., Johnston, IA (US)
Filed on Jan. 11, 2022, as Appl. No. 17/573,230.
Claims priority of provisional application 63/137,769, filed on Jan. 15, 2021.
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6806 (2018.01); C12Q 1/6874 (2018.01); C12Q 1/6895 (2018.01)
CPC C12Q 1/6895 (2013.01) [C12Q 1/6806 (2013.01); C12Q 1/6874 (2013.01); C12Q 2600/13 (2013.01)] 19 Claims
 
1. A method for genotyping a plant polynucleotide for the presence or absence of a Single Nucleotide Polymorphism (SNP) using a Surface Plasmon Resonance (SPR) assay, the method comprising the following steps:
a. Providing an oligonucleotide probe;
b. Providing a plant polynucleotide, wherein the oligonucleotide probe and plant polynucleotide are fully complementary to one another;
c. Hybridizing the oligonucleotide probe with the plant polynucleotide;
d. Complexing the oligonucleotide probe and plant polynucleotide to an SPR biosensor chip;
e. Quantitating the hybridization of the oligonucleotide probe with the plant polynucleotide to determine the affinity of oligonucleotide probe to the plant polynucleotide; and
f. Determining that the plant polynucleotide contains the presence or absence of an SNP.