	US 12,234,451 B2
	Tagmentation using immobilized transposomes with linkers
Grace Desantis, San Diego, CA (US); Stephen M. Gross, San Diego, CA (US); Jian-Sen Li, San Diego, CA (US); Natalie Morrell, Cambridge (GB); Andrew Slatter, Cambridge (GB); Kevin Shen, San Diego, CA (US); and Samantha Snow, San Diego, CA (US)
Assigned to Illumina, Inc., San Diego, CA (US)
Filed by Illumina, Inc., San Diego, CA (US); and Illumina Cambridge Limited, Cambridge (GB)
Filed on Jun. 1, 2023, as Appl. No. 18/327,187.
Application 18/327,187 is a division of application No. 17/140,434, filed on Jan. 4, 2021, granted, now 11,708,573.
Application 17/140,434 is a continuation of application No. 15/900,717, filed on Feb. 20, 2018, granted, now 10,920,219, issued on Feb. 16, 2021.
Claims priority of provisional application 62/461,620, filed on Feb. 21, 2017.
Prior Publication US 2023/0392142 A1, Dec. 7, 2023
Int. Cl. C12N 15/10 (2006.01); C12N 11/06 (2006.01); C12Q 1/6806 (2018.01); C12Q 1/6869 (2018.01)

CPC C12N 15/1065 (2013.01) [C12N 11/06 (2013.01); C12Q 1/6806 (2013.01); C12Q 1/6869 (2013.01); C12Y 207/07 (2013.01)]

21 Claims

1. A method for generating a library of tagged nucleic acid fragments from a double-stranded, target nucleic acid, comprising incubating the target nucleic acid with a transposome complex, wherein the transposome complex comprises:

a. a transposase,

b. a first transposon comprising:

i. a 3′ portion comprising a first transposon end sequence; and

ii. a first adaptor sequence at the 5′ end of the first transposon end sequence;

c. a second transposon comprising a second transposon end sequence complementary to at least a portion of the first transposon end sequence; and

d. a non-nucleic acid linker having a first end attached to the 3′ end of the second transposon and a second end attached to an affinity element,

under conditions sufficient to fragment the target nucleic acid into a plurality of target fragments, and to join the 3′ end of the first transposon to the 5′ ends of the target fragments to produce a plurality of 5′ tagged target fragments.