US 11,952,617 B2
Methods for multiplex detection of polynucleotides using unbound fluorescent probes and quencher oligonucleotides
Kiran Madanahally Divakar, Alpharetta, GA (US); and Shashi Bala, Alpharetta, GA (US)
Assigned to KASA BIO, L.L.C., Peachtree Corners, GA (US)
Filed by KASA BIO, L.L.C., Peachtree Corners, GA (US)
Filed on Mar. 17, 2023, as Appl. No. 18/186,015.
Application 18/186,015 is a continuation in part of application No. PCT/US2022/075261, filed on Aug. 22, 2022.
Claims priority of provisional application 63/235,522, filed on Aug. 20, 2021.
Prior Publication US 2023/0265490 A1, Aug. 24, 2023
Int. Cl. C12P 19/34 (2006.01); C12Q 1/6818 (2018.01); C12Q 1/6851 (2018.01)
CPC C12Q 1/6818 (2013.01) [C12Q 1/6851 (2013.01); C12Q 2600/16 (2013.01); C12Q 2600/178 (2013.01)] 30 Claims
OG exemplary drawing
 
1. A method for detection of two or more distinct target polynucleotides in a sample the method comprising
performing one or more amplification cycles on the sample using a polymerase and universal or target-specific forward and reverse primers to generate amplified products corresponding to the target polynucleotides, and
detecting and/or quantifying each amplified product using a plurality of distinct unbound detection probes and a plurality of corresponding quencher oligonucleotides that can hybridize thereto,
wherein the detecting and/or quantifying comprises measuring unbound or free detection probes.