US 12,275,984 B2
Sequential hybridization and quenching
Felice Alessio Bava, Rome (IT)
Assigned to 10X GENOMICS, INC., Pleasanton, CA (US)
Filed by 10x Genomics, Inc., Pleasanton, CA (US)
Filed on Mar. 1, 2022, as Appl. No. 17/683,856.
Claims priority of provisional application 63/155,684, filed on Mar. 2, 2021.
Prior Publication US 2022/0282306 A1, Sep. 8, 2022
Int. Cl. C12Q 1/6816 (2018.01)
CPC C12Q 1/6816 (2013.01) 20 Claims
 
1. A method for analyzing a biological sample, comprising:
a) contacting a biological sample with a first probe, wherein:
the first probe comprises a detectable moiety D1 and a hybridization sequence H1,
the biological sample comprises a nucleic acid molecule comprising a hybridization sequence H1′, and
H1′ hybridizes to H1; and
b) contacting the biological sample with a second probe, wherein:
the second probe comprises: (i) a hybridization sequence H2 that hybridizes to a hybridization region H2′ of the nucleic acid molecule, the first probe, or a splint, (ii) a quencher Q1, and (iii) a detectable moiety D2, and
upon hybridization of H2 and H2′, Q1 is in proximity to D1, thereby quenching a detectable signal from D1,
wherein a detectable signal from D2 is detected in the biological sample.