	US 12,146,133 B2
	Biotin-streptavidin cleavage composition and library fragment cleavage
Dan Cao, East Palo Alto, CA (US); Jeffrey S. Fisher, San Diego, CA (US); Fiona Kaper, Solana Beach, CA (US); Tarun Kumar Khurana, Fremont, CA (US); Tong Liu, Belmont, CA (US); Burak Okumus, San Diego, CA (US); Victor J. Quijano, Mountain View, CA (US); Clifford Lee Wang, Berkeley, CA (US); Yir-Shyuan Wu, Albany, CA (US); Shi Min Xiao, Castro Valley, CA (US); and Hongxia Xu, Castro Valley, CA (US)
Assigned to Illumina, Inc., San Diego, CA (US)
Filed by ILLUMINA, INC., San Diego, CA (US)
Filed on Feb. 3, 2021, as Appl. No. 17/166,906.
Application 17/166,906 is a continuation of application No. PCT/US2021/016228, filed on Feb. 2, 2021.
Prior Publication US 2021/0238589 A1, Aug. 5, 2021
Int. Cl. C12N 15/10 (2006.01)

CPC C12N 15/1068 (2013.01)

11 Claims

1. A method, comprising:

introducing library fragments to a flow cell, wherein the library fragments are attached to streptavidin coated solid supports;

introducing a biotin-streptavidin cleavage composition to the flow cell, the biotin-streptavidin cleavage composition including:

from about 10% by volume to about 50% by volume of a formamide reagent, the formamide reagent including 100% formamide or a combination of formamide and a buffer; and

a balance of a salt buffer; and

allowing the biotin-streptavidin cleavage composition to incubate in the flow cell at a temperature ranging from about 60° C. to about 70° C., thereby causing at least some of the library fragments to release from the solid supports and to seed to amplification primers on a surface of the flow cell.