IP Library › Granted Patent US 12,522,868
Granted Patent B2
US 12,522,868 · App. 18/341,544 · Granted Jan 13, 2026

Polynucleotides, reagents, and methods for nucleic acid hybridization

Inventors: Ramsey Ibrahim Zeitoun (Westminster, CO); Siyuan Chen (San Mateo, CA); Richard Gantt (San Francisco, CA); Kristin D. Butcher (South San Francisco, CA); E. Hutson Chilton (South San Francisco, CA)
Assignee: Twist Bioscience Corporation
C12Q1/6869C40B30/10C40B70/00G16B40/10
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Quick Facts
Patent No.
US 12,522,868
App. No.
18/341,544
Granted
Jan 13, 2026
Kind
B2
Abstract

Provided herein are compositions, methods and systems relating to libraries of polynucleotides such that the libraries allow for accurate and efficient hybridization after binding to target sequences. Further provided herein are probes, blockers, additives, buffers, and methods that result in improved hybridization. Such compositions and methods are useful for improvement of Next Generation Sequencing applications, such as reducing off-target binding or reducing workflow times.

Claims (21)

1 . A method for nucleic acid hybridization, comprising:

(a) contacting a first polynucleotide library with a second polynucleotide library, wherein at least one polynucleotide in the first polynucleotide library comprises a target sequence, at least one polynucleotide in the second polynucleotide library comprises a target binding sequence that is complementary to the target sequence, and wherein at least one and less than or equal to 90% of polynucleotides in the second polynucleotide library comprise a label;

(b) enriching the at least one polynucleotide comprising the target sequence library to generate at least one enriched target polynucleotide; and

(c) sequencing the at least one enriched target polynucleotide.

2 . The method of claim 1 , wherein less than or equal to 50% of the polynucleotides in the second polynucleotide library comprise the label.

3 . The method of claim 1 , wherein less than or equal to 30% of the polynucleotides in the second polynucleotide library comprise the label.

4 . The method of claim 1 , wherein less than or equal to 15% of the polynucleotides in the second polynucleotide library comprise the label.

5 . The method of claim 1 , wherein the label comprises a molecular tag.

6 . The method of claim 1 , wherein the label comprises biotin.

7 . The method of claim 1 , wherein contacting the first polynucleotide library with the second polynucleotide library decreases an AT dropout rate or a GC dropout rate.

8 . The method of claim 7 , wherein the AT dropout rate or the GC dropout rate is less than 2%.

9 . The method of claim 1 , wherein contacting the first polynucleotide library with the second polynucleotide library decreases a percent off bait rate.

10 . The method of claim 9 , wherein the percent off bait rate is less than 25%.

11 . The method of claim 1 , wherein the first polynucleotide library comprises fragments of genomic DNA.

12 . The method of claim 1 , wherein the first polynucleotide library comprises an exome library.

13 . The method of claim 1 , wherein the second polynucleotide library comprises an exome panel and a gene panel targeting one or more polynucleotides of the first polynucleotide library corresponding to fragments of genomic DNA.

14 . The method of claim 1 , wherein the second polynucleotide library comprises two gene panels targeting one or more polynucleotides of the first polynucleotide library corresponding to fragments of genomic DNA.

15 . The method of claim 1 , wherein a number of polynucleotides captured in the first polynucleotide library in (a) varies depending on a percent of labeled polynucleotides of the second polynucleotide library.

16 . The method of claim 1 , wherein sequencing the at least one enriched target polynucleotide comprises sequencing the at least one enriched target polynucleotide such that at least 95% of base pairs within the at least one enriched target polynucleotide have a read depth less than or equal to a mean read depth.

17 . The method of claim 1 , wherein sequencing the at least one enriched target polynucleotide comprises sequencing the at least one enriched target polynucleotide such that at least 90% of base pairs within the at least one enriched target polynucleotide have a 30× read depth.

18 . The method of claim 1 , further comprising, prior to (a), providing the first polynucleotide library and the second polynucleotide library, and labeling less than 90% of the polynucleotides in the second polynucleotide library, wherein a ratio of polynucleotides in the first polynucleotide library to the second polynucleotide library is constant.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 11, 2024
From: ZEITOUN, RAMSEY IBRAHIM; CHEN, SIYUAN; GANTT, RICHARD; BUTCHER, KRISTIN D.; CHILTON, E. HUTSON
To: TWIST BIOSCIENCE CORPORATION
Reel/Frame 066717/0845 →
Continuity (10)
Continuation 16590301 · Oct 1, 2019
Continuation 16417023 · May 20, 2019
Provisional Application 62833440 · Apr 12, 2019
Provisional Application 62814749 · Mar 6, 2019
Provisional Application 62814753 · Mar 6, 2019
Provisional Application 62810293 · Feb 25, 2019
Provisional Application 62810343 · Feb 25, 2019
Provisional Application 62675647 · May 23, 2018
Provisional Application 62673704 · May 18, 2018
Related Publication 20240043920A1 · Feb 8, 2024
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