	US 12,486,531 B2
	Methods for preparing nucleic acid molecules for sequencing
Wigard Pieter Kloosterman, Utrecht (NL); Jeroen de Ridder, Utrecht (NL); and Alessio Marcozzi, Utrecht (NL)
Assigned to UMC UTRECHT HOLDING B.V., Utrecht (NL)
Filed by UMC UTRECHT HOLDING B.V.
Filed on Sep. 11, 2023, as Appl. No. 18/465,025.
Application 18/465,025 is a continuation of application No. 16/771,653, granted, now 11,781,170, previously published as PCT/NL2018/050831, filed on Dec. 11, 2018.
Claims priority of application No. 17206503 (EP), filed on Dec. 11, 2017.
Prior Publication US 2024/0132937 A1, Apr. 25, 2024
Int. Cl. C12Q 1/6806 (2018.01); C12Q 1/6869 (2018.01)

CPC C12Q 1/6806 (2013.01) [C12Q 1/6869 (2013.01)]

11 Claims

1. A collection of linear backbone DNA molecules of a length of 20-1000 nucleotides that comprise 5′ ends that comprise a part of a first restriction enzyme recognition site at the extreme end and 3′ ends that comprise the other part of a first restriction enzyme recognition site at the extreme end, and which 5′ and 3′ ends are ligation compatible with each other and form a restriction enzyme recognition site when self-ligated and wherein each of said backbones comprises:

a linker;

an identifier sequence that differs from the sequence of identifiers of other backbones in the collection; and

optionally a restriction site for a nicking enzyme;

wherein:

said collection of linear backbone DNA molecules have a flexibility score of 10 or more; and

said linker comprises a sequence of 20-900 nucleotides.