US 12,264,358 B2
Method of selectively sequencing amplicons in a biological sample
George M. Church, Brookline, MA (US); Jehyuk Lee, Allston, MA (US); Richard C. Terry, Carlisle, MA (US); and Evan R. Daugharthy, Cambridge, MA (US)
Assigned to President and Fellows of Harvard College, Cambridge, MA (US)
Filed by President and Fellows of Harvard College, Cambridge, MA (US)
Filed on May 2, 2024, as Appl. No. 18/652,923.
Application 18/652,923 is a continuation of application No. 17/708,050, filed on Mar. 30, 2022.
Application 17/708,050 is a continuation of application No. 17/363,097, filed on Jun. 30, 2021, granted, now 11,299,767, issued on Apr. 12, 2022.
Application 17/363,097 is a continuation of application No. 16/157,243, filed on Oct. 11, 2018, granted, now 11,078,520, issued on Aug. 3, 2021.
Application 16/157,243 is a continuation of application No. 14/774,282, granted, now 10,138,509, issued on Nov. 27, 2018, previously published as PCT/US2014/018580, filed on Feb. 26, 2014.
Claims priority of provisional application 61/777,383, filed on Mar. 12, 2013.
Prior Publication US 2024/0287582 A1, Aug. 29, 2024
Int. Cl. C12Q 1/6806 (2018.01); C12P 19/34 (2006.01); C12Q 1/6844 (2018.01); C12Q 1/6869 (2018.01); C12Q 1/6874 (2018.01)
CPC C12Q 1/6806 (2013.01) [C12P 19/34 (2013.01); C12Q 1/6844 (2013.01); C12Q 1/6869 (2013.01); C12Q 1/6874 (2013.01)] 29 Claims
OG exemplary drawing
 
1. A method, comprising,
(a) providing a biological sample comprising a plurality of cells comprising a plurality of ribonucleic acid (RNA) molecules;
(b) generating a plurality of circularized nucleic acid molecules in said biological sample, wherein each of said plurality of circularized nucleic acid molecules comprises a nucleic acid sequence corresponding to a sequence of an RNA molecule of said plurality of RNA molecules;
(c) amplifying circularized nucleic acid molecules of said plurality of circularized nucleic acid molecules in a rolling circle amplification reaction to generate a plurality of amplicons in said biological sample; and
(d) using selective sequencing primers to sequence different subsets of said plurality of amplicons in said biological sample.