US 12,435,362 B2
Compositions and methods for improving library enrichment
Andrew F. Slatter, Cambridge (GB); Junhua Zhao, San Diego, CA (US); Grace DeSantis, San Diego, CA (US); Stephen M. Gross, San Diego, CA (US); Hayley Marianne Hogan, San Diego, CA (US); Jian-Sen Li, San Diego, CA (US); and Oliver Jon Miller, Cambridge (GB)
Assigned to Illumina, Inc., San Diego, CA (US); and Illumina Cambridge Limited, Cambridge (GB)
Appl. No. 17/267,107
Filed by Illumina, Inc., San Diego, CA (US); and Illumina Cambridge Limited, Cambridge (GB)
PCT Filed Aug. 13, 2019, PCT No. PCT/US2019/046396
§ 371(c)(1), (2) Date Feb. 9, 2021,
PCT Pub. No. WO2020/036991, PCT Pub. Date Feb. 20, 2020.
Claims priority of provisional application 62/764,753, filed on Aug. 15, 2018.
Prior Publication US 2021/0164027 A1, Jun. 3, 2021
Int. Cl. C12Q 1/6832 (2018.01); C12Q 1/6869 (2018.01)
CPC C12Q 1/6832 (2013.01) [C12Q 1/6869 (2013.01)] 16 Claims
 
1. A hybridization buffer comprising: a crowding agent, and human Cot-1 DNA, a destabilizing agent, salt, and a blocker, wherein the blocker comprises at least one of:
(a) an oligonucleotide and is capable of binding to an adaptor, wherein the adaptor comprises a universal primer sequence and at least one of an index region or a unique molecular identifier (UMI), wherein a first region of the blocker capable of binding to the index region and/or UMI of the adaptor comprises: at least three thymine bases, at least four thymine bases, at least five thymine bases, at least six thymine bases, at least seven thymine bases, at least eight thymine bases, at least nine thymine bases, or at least ten thymine bases, and wherein a second region of the blocker capable of binding to a non-index region of the adaptor comprises modified nucleotides; or
(b) two non-connected oligonucleotides; wherein the blocker is capable of binding to at least a portion of an adaptor, wherein the adaptor comprises a universal primer sequence and at least one of an index region or a unique molecular identifier (UMI); and wherein the non-connected oligonucleotides comprise bases that do not correspond to the index region and/or the UMI of the adaptor.