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US 2022/0177873 A1Published application
Isolation and immobilization of nucleic acids and uses thereof
AssigneeSimplseq, Inc.
InventorBrandon Michael YOUNG
Life of this patent
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Abstract
Disclosed is a novel technique of directly isolating nucleic acids from a biological sample and use of the isolated nucleic acid complexes for various applications and assays such as biobanking and sequencing.
Claims24
-14. (canceled)
A method of generating a cDNA library from a biological sample, comprising:
- obtaining a plurality of isolated nucleic acid fragments bound to a solid support;
- hybridizing one or more primers to the plurality of the isolated nucleic acid fragments;
- conducting primer extension using the plurality of the isolated nucleic acid fragments as templates to synthesize cDNA fragments;
- and
- eluting the synthesized cDNA fragments from the solid support to generate the cDNA library,
- wherein the plurality of the isolated nucleic acid fragments is obtained by contacting a biological sample containing the plurality of the nucleic acid fragments with a reaction mix comprising a template-independent DNA or RNA polymerase, and one or more NTPs, dNTPs or ddNTPs to obtain a tagged nucleic acid, wherein at least a fraction of the one or more NTPs, dNTPs or ddNTPs are modified by an affinity tag;
- binding the tagged nucleic acid fragments to a surface of a solid support;
- and
- washing the solid support to obtain the bound nucleic acid fragments isolated from the biological sample.
6 more elements
A captured nucleic acid complex comprising:
- a solid support;
- and
- a plurality of nucleic acid fragments, each fragment having a 3′ end bound to a polymeric tail, and each polymeric tail is bound to a surface of the solid support.
25-31. (canceled)
Dependencies are inferred from the claim text.