Compositions, methods, systems and kits for target nucleic acid enrichment
AssigneeLife Technologies Corporation
InventorsMark Andersen, Steven Roman
Life of this patent
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Abstract
The present invention provides methods, compositions, kits, systems and apparatus that are useful for isolating nucleic acid molecules from a sample. In particular, the methods generally relate to normalizing the concentration of target nucleic acid molecules from a sample. In one aspect, the invention relates to purifying a primer extension product from a primer extension reaction mixture. In some aspects, nucleic acid molecules obtained using the disclosed methods, kits, systems and apparatuses can be used in various downstream processes including nucleic acid sequencing.
Claims20
A method for normalizing a desired amount of nucleic acid from a sample, comprising:
- a) generating a population of bound target nucleic acid molecules by contacting a sample including a population of target nucleic acid molecules with a hairpin primer which include a first primer sequence and a second primer sequence separated by a non-replicable moiety, wherein the second primer sequence binds a portion of the target nucleic acid molecules, and wherein the first primer sequence binds a capture oligonucleotide, under conditions where at least some of the hairpin primers hybridize to at least some of the target nucleic acid molecules;
- and
- b) forming a population of captured target nucleic acid molecules by contacting the bound target nucleic acid molecules with a limiting, desired amount of the capture oligonucleotides, wherein the capture oligonucleotide includes a capture moiety, thereby capturing a desired portion of the population of bound target nucleic acid molecules, wherein the number of captured target nucleic acid molecules is directly proportional to the limiting number of capture oligonucleotides.
A method for normalizing a desired amount of nucleic acid from a sample, comprising:
- a) generating a population of bound target nucleic acid molecules by contacting a sample including a population of target nucleic acid molecules with a hairpin primer which include a first primer sequence and a second primer sequence separated by a non-replicable moiety, wherein the second primer sequence binds a portion of the target nucleic acid molecules, and wherein the first primer sequence binds a capture oligonucleotide, under conditions where at least some of the hairpin primers hybridize to at least some of the target nucleic acid molecules;
- b) forming a population of captured target nucleic acid molecules by contacting the bound target nucleic acid molecules with a limiting, desired amount of the capture oligonucleotides and a binding agent, wherein the capture oligonucleotide includes a capture moiety and the capture moiety binds the binding agent, thereby capturing a desired portion of the population of bound target nucleic acid molecules, wherein the number of captured target nucleic acid molecules is directly proportional to the limiting number of capture oligonucleotides;
- and
- c) subjecting the captured nucleic acid molecules to non-permissive conditions to elute the captured nucleic acid molecules from the binding agent, thereby forming a normalized population of target nucleic acid molecules.
Dependencies are inferred from the claim text.