US 12,331,356 B2
Multipart reagents having increased avidity for polymerase binding
Sinan Arslan, San Diego, CA (US); Chunhong Zhou, San Diego, CA (US); Molly Min He, San Diego, CA (US); Matthew Kellinger, San Diego, CA (US); Hui Zhen Mah, San Diego, CA (US); Michael Previte, San Diego, CA (US); and Lei Sun, San Diego, CA (US)
Assigned to ELEMENT BIOSCIENCES, INC., San Diego, CA (US)
Filed by ELEMENT BIOSCIENCES, INC., San Diego, CA (US)
Filed on Jan. 22, 2021, as Appl. No. 17/156,452.
Application 17/156,452 is a continuation of application No. 16/855,877, filed on Apr. 22, 2020, granted, now 10,982,280.
Application 16/855,877 is a continuation of application No. 16/740,357, filed on Jan. 10, 2020, granted, now 10,704,094.
Application 16/855,877 is a continuation of application No. PCT/US2019/061556, filed on Nov. 14, 2019.
Application 16/740,357 is a continuation in part of application No. 16/579,794, filed on Sep. 23, 2019, granted, now 10,768,173.
Application PCT/US2019/061556 is a continuation in part of application No. 16/363,842, filed on Mar. 25, 2019, abandoned.
Claims priority of provisional application 62/897,172, filed on Sep. 6, 2019.
Claims priority of provisional application 62/776,898, filed on Dec. 7, 2018.
Claims priority of provisional application 62/767,343, filed on Nov. 14, 2018.
Prior Publication US 2021/0139981 A1, May 13, 2021
Int. Cl. C12Q 1/6876 (2018.01); C12N 9/12 (2006.01); G01N 21/64 (2006.01); G01N 33/543 (2006.01)
CPC C12Q 1/6876 (2013.01) [C12N 9/1241 (2013.01); G01N 21/6428 (2013.01); G01N 33/54306 (2013.01); G01N 2021/6439 (2013.01)] 20 Claims
 
1. A method for nucleic acid identification, the method comprising:
a) providing a surface having coupled thereto at least one polymer layer comprising a hydrophilic polymer coupled to a first nucleic acid molecule;
b) introducing a fluid composition comprising a plurality of nucleic acid molecules comprising a second nucleic acid molecule at a concentration of less than 1 nanomolar to said surface under conditions sufficient to hybridize at least a portion of said second nucleic acid molecule to at least a portion of said first nucleic acid molecule while reducing a nonspecific coupling of non-hybridized nucleic acids of said plurality of nucleic acid molecules to said surface as compared to a method performed without said surface or under said conditions, wherein said second nucleic acid molecule provides a template for a nucleotide binding reaction;
c) contacting said second nucleic acid molecule or derivative thereof with a detectable nucleotide moiety under conditions sufficient to cause the nucleotide binding reaction to occur between said detectable nucleotide moiety and a nucleotide of said second nucleic acid molecule or derivative thereof; and
d) detecting said detectable nucleotide moiety bound to said nucleotide of said second nucleic acid molecule or derivative thereof, thereby identifying said nucleotide,
wherein said method uses a reduced concentration of said second nucleic acid molecule and results in a shortened hybridization time as compared to a method that is not performed using said surface under said conditions.