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US 11,905,553 B2Granted
Method for nucleic acid amplification
AssigneeSt. Jude Children's Research Hospital
InventorsCharles Gawad, John Easton, Veronica Gonzalez-Pena
Life of this patent
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Abstract
Provided herein are compositions and methods for accurate and scalable Primary Template-Directed Amplification (PTA) nucleic acid amplification and sequencing methods, and their applications for research, diagnostics, and treatment.
Claims20
A method of amplifying at least one target nucleic acid molecule, the method comprising:
- a. contacting a sample comprising the at least one target nucleic acid molecule, at least one amplification primer, at least one nucleic acid polymerase comprising 3′-5′ exonuclease activity, and a mixture of nucleotides, wherein the mixture of nucleotides comprises at least one terminator nucleotide which terminates nucleic acid replication by the polymerase, wherein the terminator nucleotide is an irreversible terminator which inhibits 3′-5′ exonuclease activity of the polymerase, and is selected from nucleotides with modification to the alpha group, C3 spacer nucleotides, locked nucleic acids (LNA), inverted nucleic acids, 2′ fluoro nucleotides, 3′ phosphorylated nucleotides, 2′-O-Methyl modified nucleotides, and trans nucleic acids, and
- b. amplifying the at least one target nucleic acid molecule to generate a plurality of terminated amplification products, wherein the terminator nucleotide is attached to the 3′ terminus of the terminated amplification products, and wherein the replication proceeds by strand displacement replication.
Dependencies are inferred from the claim text.