Polynucleotides, reagents, and methods for nucleic acid hybridization
AssigneeTwist Bioscience Corporation
InventorsRamsey Ibrahim ZEITOUN, Siyuan CHEN, Richard GANTT, Kristin D. BUTCHER, E. Hutson CHILTON
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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.
Claims21
1-107. (canceled)
A composition for nucleic acid hybridization comprising:
- a first polynucleotide library wherein each polynucleotide in the first library comprises at least one index sequence, and at least one adapter sequence, and wherein at least one polynucleotide in the first polynucleotide library comprises a target sequence;
- a second polynucleotide library, wherein at least one polynucleotide in the second polynucleotide library comprises a target binding sequence that is complementary to the target sequence;
- and
- at least one polynucleotide blocker, wherein the polynucleotide blocker is complementary to at least a portion of the adapter sequence, but not complementary to the at least one index sequence.
A method for reducing off-target nucleic acid hybridization, comprising:
- contacting a first polynucleotide library, a second polynucleotide library, and at least one polynucleotide blocker, wherein:
- at least one polynucleotide in the first polynucleotide library comprises a target sequence,
- each polynucleotide in the first polynucleotide library comprises at least one index sequence, and at least one adapter sequence,
- at least one polynucleotide in the second polynucleotide library comprises a target binding sequence that is complementary to the target sequence of the at least one polynucleotide in the first polynucleotide library, and
- the at least one polynucleotide blocker is complementary to at least a portion of the adapter sequence, but not complementary to the at least one index sequence;
- enriching the at least one polynucleotide comprising the target sequence to generate at least one enriched target polynucleotide;
- and
- sequencing the at least one enriched target polynucleotide.
Dependencies are inferred from the claim text.