USOO933.4531B2 (12) United States Patent (10) Patent No.: US 9,334,531 B2 Li et al. (45) Date of Patent: *May 10, 2016 (54) NUCLECACIDAMPLIFICATION (56) References Cited U.S. PATENT DOCUMENTS (71) Applicant: LIFE TECHNOLOGIES 5,223,414 A 6/1993 Zarling et al. CORPORATION, Carlsbad, CA (US) 5,616,478 A 4/1997 Chetverin et al. 5,670,325 A 9/1997 Lapidus et al. (72) Inventors: Chieh-Yuan Li, El Cerrito, CA (US); 5,928,870 A 7/1999 Lapidus et al. David Ruff, San Francisco, CA (US); 5,958,698 A 9, 1999 Chetverinet al. Shiaw-Min Chen, Fremont, CA (US); 6,001,568 A 12/1999 Chetverinet al. 6,033,881 A 3/2000 Himmler et al. Jennifer O'Neil, Wakefield, MA (US); 6,074,853. A 6/2000 Pati et al. Rachel Kasinskas, Amesbury, MA (US); 6,306,590 B1 10/2001 Mehta et al. Jonathan Rothberg, Guilford, CT (US); 6.432,360 B1 8, 2002 Church Bin Li, Palo Alto, CA (US); Kai Qin 6,440,706 B1 8/2002 Vogelstein et al. Lao, Pleasanton, CA (US) 6,511,803 B1 1/2003 Church et al. 6,929,915 B2 8, 2005 Benkovic et al. 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US 2014/O080717 A1 Mar. 20, 2014 8,062,850 B2 11/2011 Piepenburg et al. Related U.S. Application Data (Continued) (63) Continuation-in-part of application No. 13/923,232, FOREIGN PATENT DOCUMENTS filed on Jun. 20, 13, which is a continuation of CN 1489632 A2 4/2004 application No. 13/842.296, filed on Mar. 15, 2013, CN 1014 13034 B 2, 2011 which is a continuation-in-part of application No. (Continued) 13/328,844, filed on Dec. 16, 2011, now abandoned, OTHER PUBLICATIONS said application No. 13/842.296 is a Bentley et al. (Accurate Whole Human Genome Sequencing using continuation-in-part of application No. 13/828,049, Reversible Terminator Chemistry, Nature. Nov. 6, 2008; 4.56(7218): filed on Mar. 14, 2013, which is a continuation-in-part 53-59).* of application No. 13/328,844, filed on Dec. 16, 2011, now abandoned, said application No. 13/842.296 is a (Continued) continuation-in-part of application No. Primary Examiner — Aaron Priest PCT/US2011/065535, filed on Dec. 16, 2011, application No. 14/023,361, which is a (57) ABSTRACT continuation-in-part of application No. In some embodiments, the present teachings provide methods PCT/US2013/037352, filed on Apr. 19, 2013. for nucleic acid amplification, comprising forming a reaction mixture, and Subjecting the reaction mixture to conditions (60) Provisional application No. 61/876,136, filed on Sep. Suitable for nucleic acid amplification. In some embodiments, 10, 2013, provisional application No. 61/859,000, methods for nucleic acid amplification include Subjecting the filed on Jul. 26, 2013, provisional application No. nucleic acid to be amplified to partially denaturing condi 61/858,977, filed on Jul. 26, 2013, provisional tions. In some embodiments, methods for nucleic acid ampli application No. 61/822,239, filed on May 10, 2013, fication include amplifying without fully denaturing the provisional application No. 61/822.226, filed on May nucleic acid that is amplified. In some embodiments, the 10, 2013, provisional application No. 61/792.247, methods for nucleic acid amplification employ an enzyme filed on Mar. 15, 2013, provisional application No. that catalyzes homologous recombination and a polymerase. In some embodiments, methods for nucleic acid amplifica (Continued) tion can be conducted in a single reaction vessel. In some embodiments, methods for nucleic acid amplification can be (51) Int. C. conducted in a single continuous liquid phase of a reaction CI2O I/68 (2006.01) mixture, without need for compartmentalization of the reac (52) U.S. C. tion mixture or immobilization of reaction components. In CPC ............ CI2O I/6853 (2013.01): CI2O I/6846 Some embodiments, methods for nucleic acid amplification (2013.01); C12O 1/6855 (2013.01); C12O comprise a amplifying at least one polynucleotide onto a I/6874 (2013.01) Surface under isothermal amplification conditions, optionally (58) Field of Classification Search in the presence of a polymer. The polymer can include a None sieving agent and/or a diffusion-reducing agent. 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