US 12,435,370 B2
Methods and compositions for sequencing double stranded nucleic acids
Kurt Patterson, San Diego, CA (US); Hari K. K. Subramanian, San Diego, CA (US); Brittany A. Rohrman, San Diego, CA (US); and Fabian Block, San Diego, CA (US)
Assigned to Pacific Biosciences of California, Inc., Menlo Park, CA (US)
Filed by Pacific Biosciences of California, Inc., Menlo Park, CA (US)
Filed on Mar. 16, 2023, as Appl. No. 18/184,928.
Application 18/184,928 is a continuation of application No. 17/814,062, filed on Jul. 21, 2022, granted, now 11,608,528.
Application 17/814,062 is a continuation of application No. PCT/US2021/020564, filed on Mar. 3, 2021.
Claims priority of provisional application 62/984,438, filed on Mar. 3, 2020.
Prior Publication US 2023/0357844 A1, Nov. 9, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12P 19/34 (2006.01); C12Q 1/6874 (2018.01); C12Q 1/6806 (2018.01); C12Q 1/6853 (2018.01)
CPC C12Q 1/6874 (2013.01) [C12Q 1/6806 (2013.01); C12Q 1/6853 (2013.01)] 19 Claims
OG exemplary drawing
 
1. A method for determining sequences from sense and antisense strands of a nucleic acid, comprising
(a) providing a nucleic acid cluster attached to a solid support, wherein the nucleic acid cluster comprises a sense strand of a concatemer and an antisense strand of the concatemer, wherein the concatemer comprises multiple copies of a sequence unit linked in series, wherein the sequence unit comprises a target sequence and a primer binding site;
(b) hybridizing a primer to the primer binding site in a sequence unit of the antisense strand in the cluster;
(c) extending the primer along the antisense strand to determine the sequence from at least a portion of the target sequence in the antisense strand;
(d) removing the antisense strand from the cluster prior to steps (e) and (f);
(e) hybridizing a second primer to the primer binding site in a sequence unit of the sense strand; and
(f) extending the second primer along the sense strand to determine the sequence from at least a portion of the target sequence in the sense strand.