	US 11,898,198 B2
	Universal short adapters with variable length non-random unique molecular identifiers
Chen Zhao, San Diego, CA (US); Kevin Wu, San Diego, CA (US); Han-Yu Chuang, San Diego, CA (US); Jennifer Lococo, San Diego, CA (US); Alex So, San Diego, CA (US); Dwight Baker, San Diego, CA (US); and Tatjana Singer, San Diego, CA (US)
Assigned to Illumina, Inc., San Diego, CA (US)
Filed by Illumina, Inc., San Diego, CA (US)
Filed on Aug. 17, 2022, as Appl. No. 17/890,208.
Application 17/890,208 is a continuation of application No. 16/129,099, filed on Sep. 12, 2018, granted, now 11,447,818.
Claims priority of provisional application 62/559,448, filed on Sep. 15, 2017.
Prior Publication US 2023/0242977 A1, Aug. 3, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12P 19/34 (2006.01); C12Q 1/6855 (2018.01); G16B 20/20 (2019.01); G16B 35/10 (2019.01); G16B 30/10 (2019.01); G16B 25/20 (2019.01); G16B 40/00 (2019.01); G16B 20/00 (2019.01); G16B 30/00 (2019.01); C12Q 1/686 (2018.01); C12Q 1/6869 (2018.01); C12Q 1/6876 (2018.01)

CPC C12Q 1/6855 (2013.01) [C12Q 1/686 (2013.01); C12Q 1/6869 (2013.01); C12Q 1/6876 (2013.01); G16B 20/00 (2019.02); G16B 20/20 (2019.02); G16B 25/20 (2019.02); G16B 30/00 (2019.02); G16B 30/10 (2019.02); G16B 35/10 (2019.02); G16B 40/00 (2019.02); C12Q 2525/191 (2013.01); C12Q 2525/197 (2013.01); C12Q 2600/16 (2013.01); C12Q 2600/166 (2013.01)]

20 Claims

1. A set of sequencing adapters comprising a plurality of double-stranded polynucleotides, wherein:

each double-stranded polynucleotide comprises a double-stranded hybridized region, a single-stranded 5′ arm, a single-stranded 3′ arm, and at least one variable-length, nonrandom unique molecular index (vNRUMI);

variable-length, nonrandom unique molecular indices (vNRUMIs) of the set of sequencing adapters form a set of vNRUMIs configured to identify individual nucleic acid molecules in a sample for multiplex massively parallel sequencing;

the set of vNRUMIs comprises sequences having two or more molecular lengths;

an edit distance between any two vNRUMIs of the set of vNRUMIs is not less than a first criterion value, wherein the first criterion value is at least two; and

edit distances among the vNRUMIs in the set of vNRUMIs are maximized by a greedy approach.