Polynucleotides, reagents, and methods for nucleic acid hybridization

Patent metadata

Field	Value
Publication number	US-11732294-B2
Application number	US-201916590301-A
Country	US
Kind code	B2
Filing date	Oct 1, 2019
Priority date	May 18, 2018
Publication date	Aug 22, 2023
Grant date	Aug 22, 2023

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.

First claim

What is claimed is:

1. An aqueous composition for polynucleotide hybridization comprising:
at least one solvent, wherein the at least one solvent comprises dimethyl sulfoxide (DMSO) at 10-30% (v/v);
at least one surfactant; and
at least one thickening agent.

2. The composition of claim 1 , wherein the at least one solvent further comprises DMF, DMAc, or HEMPA.

3. The composition of claim 1 , wherein the at least one solvent further comprises acetone, methyl ethyl ketone, THF, or diethyl ether.

4. The composition of claim 1 , wherein the at least one surfactant comprises SDS, CTAB, tergitol, or sodium lauryl sulfate.

5. The composition of claim 1 , wherein the at least one surfactant comprises a trialkylammonium salt.

6. The composition of claim 1 , wherein the at least one thickening agent comprises cellulose, or starch.

7. The composition of claim 1 , wherein the at least one thickening agent comprises dextran sulfate, hydroxymethylcellulose, hydroxyethylcellulose, polyethylene glycol, or ficoll.

8. The composition of claim 1 , wherein the at least one thickening agent is 10-40% (w/v).

9. The composition of claim 1 , wherein the at least one surfactant is 0.001-0.1% (w/v).

10. The composition of claim 4 , wherein the at least one surfactant is 0.001-0.1% (w/v).

11. The composition of claim 5 , wherein the at least one surfactant is 0.001-0.1% (w/v).

12. The composition of claim 6 , wherein the at least one thickening agent is 10-40% (w/v).

13. The composition of claim 7 , wherein the at least one thickening agent is 10-40% (w/v).

14. The composition of claim 1 , wherein a hybridization temperature of the composition is at least 50° C.

15. The composition of claim 1 , wherein a hybridization temperature of the composition is at least 65° C.

16. The composition of claim 1 , wherein the aqueous composition comprises a plurality of polynucleotides.

17. The composition of claim 16 , wherein the plurality of polynucleotides are in solution.

Assignees

• Twist Bioscience Corp

Inventors

• ZEITOUN Ramsey Ibrahim
• Chen Siyuan
• Gantt Richard
• Butcher Kristin D
• Chilton E Hutson

Classifications

• C40B40/06

  Libraries containing nucleotides or polynucleotides, or derivatives thereof · CPC title

• C12Q1/6827 Primary

  for detection of mutation or polymorphism · CPC title

• C12Q2600/156

  Polymorphic or mutational markers · CPC title

• C40B50/14 Primary

  Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support · CPC title

• C12Q1/6832

  Enhancement of hybridisation reaction · CPC title