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US 11,268,087 B2Granted
Isolation and immobilization of nucleic acids and uses thereof
AssigneeSimplseq, Inc.
InventorBrandon Michael Young
Life of this patent
Estimating the term…
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.
Claims29
A method of isolating nucleic acid fragments, comprising:
- contacting a biological sample containing nucleic acid fragments with a reaction mix comprising a template-independent DNA or RNA polymerase, and one or more NTPs, dNTPs or ddNTPs, wherein at least a fraction of the one or more NTPs, dNTPs or ddNTPs are modified by an affinity tag, to obtain tagged nucleic acid fragments;
- 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,
- wherein the biological sample is selected from the group consisting of blood, plasma, serum, urine, saliva, exosome, a lysed formalin-fixed, paraffin-embedded (FFPE) tissue sample, and cells.
2 more elements
A method of isolating nucleic acid fragments comprising:
- combining a blood or plasma sample containing nucleic acid fragments with a reaction mix comprising a template-independent DNA or RNA polymerase, and one or more NTPs, dNTPs or ddNTPs, wherein at least a fraction of the one or more NTPs, dNTPs or ddNTPs are modified by a biotin affinity tag, to obtain nucleic acid fragments having a biotin tagged polymeric tail;
- binding the biotin tagged nucleic acid fragments to a solid support surface coated with avidin, streptavidin, or neutravidin;
- and
- washing the solid support to isolate the bound nucleic acid fragments from the blood or plasma sample.
1 more element
A method of isolating DNA fragments comprising:
- combining a blood or plasma sample containing DNA fragments with a reaction mix comprising a template-independent DNA polymerase, and one or more NTPs, dNTPs or ddNTPs, wherein at least a fraction of the NTPs, dNTPs or ddNTPs are modified by a biotin affinity tag, to obtain nucleic acid fragments having a biotin tagged polymeric tail;
- binding the biotin tagged DNA fragments to a solid support surface coated with avidin, streptavidin, or neutravidin;
- and
- washing the solid support to isolate the bound DNA fragments from the blood or plasma sample.
1 more element
A method of isolating DNA fragments comprising:
- denaturing double stranded DNA fragments in a blood or plasma sample into single stranded DNA fragments having a 5′ end and a 3′ end;
- combining the blood or plasma sample containing DNA fragments with a reaction mix comprising a terminal transferase (TdT), and one or more NTPs, dNTPs or ddNTPs, wherein at least a fraction of the NTPs, dNTPs or ddNTPs are modified by a biotin affinity tag, to obtain DNA fragments having a biotin tagged polymeric tail at the 3′ end;
- binding the biotin tagged DNA fragments to a solid support surface coated with avidin, streptavidin, or neutravidin;
- and
- washing the solid support to isolate the bound DNA fragments from the blood or plasma sample.
2 more elements
Dependencies are inferred from the claim text.