Process for labeling a nucleic acid
AssigneesBio Merieux, Gen-Probe Incorporated
InventorsAli Laayoun, Lionel Menou, Christelle Tora, Aloke R. Banerjee, Michael M. Becker, Kenneth A. Browne, Matthew C. Friedenberg, Fred F. Hajjar
Life of this patent
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Abstract
A process of fragmenting and labeling a synthetic or natural nucleic acid, comprising the steps of providing a mixture containing a nucleic acid, a labeling agent containing a detectable label, and at least one multivalent metal cation in a substantially aqueous solution; chemically fragmenting the nucleic acid in the mixture to produce a multiplicity of nucleic acid fragments; and attaching at least one label to at least one of the nucleic acid fragments to produce a detectably labeled nucleic acid fragment.
Claims49
A process for fragmenting and labeling at least one synthetic or natural DNA, RNA or chimeric DNA-RNA polymer, comprising the steps of:
- obtaining a mixture in which the at least one DNA, RNA or chimeric DNA-RNA polymer has been subject to an in vitro nucleic acid amplification reaction;
- chemically fragmenting the at least one DNA, RNA or chimeric DNA RNA polymer in the mixture in the presence of at least one multivalent metal cation in an aqueous solution, to produce a plurality of DNA or RNA fragments having freed terminal phosphates for further reaction;
- and
- attaching a labeling agent on a plurality of said fragments at freed terminal phosphates located at the 3′ end and/or 5′ end of said fragments, wherein the fragmenting and attaching steps take place in an in vitro nucleic acid amplification mixture.
A process for fragmenting and labeling a synthetic or natural DNA or RNA nucleic acid, comprising the steps of:
- obtaining a mixture in which the DNA or RNA nucleic acid has been subject to an in vitro nucleic acid amplification reaction;
- chemically fragmenting said nucleic acid in the mixture in the presence of at least one multivalent metal cation in an aqueous solution, to produce a plurality of DNA or RNA fragments having freed terminal phosphates for further reaction;
- attaching a labeling agent on a plurality of said fragments at freed terminal phosphates located at the 3′ end and/or 5′ end of said fragments;
- and
- treating said mixture to decrease or eliminate unattached labeling agent, wherein the treating step physically separates the labeled nucleic acid fragment from unattached labeling agent in the mixture after the fragmenting and attaching steps, and wherein the treating step dilutes the mixture.
The process according to claim wherein the fragmenting step for RNA or chimeric DNA-RNA polymer is performed in the presence of at least one multivalent metal cation selected from the group consisting of Cr3+, Ce3+, Yb3+, Tb3+, Eu2+ and Pb2+.
Dependencies are inferred from the claim text.