Methods and compositions for analyzing nucleic acid molecules utilizing sizing techniques
AssigneeQIAGEN Genomics, Inc.
InventorsJeffrey Van Ness, John C. Tabone, J. Jeffry Howbert, John T. Mulligan
Life of this patent
Estimating the term…
Abstract
Tags and linkers specifically designed for a wide variety of nucleic acid reactions are disclosed, which are suitable for a wide variety of nucleic acid reactions wherein separation of nucleic acid molecules based upon size is required.
Claims40
A method for determining the identity of a nucleic acid molecule, comprising sequentially:
- (a) generating tagged nucleic acid molecules from one or more selected target nucleic acid molecules, wherein a tag is correlative with a particular nucleic acid fragment and detectable by non-fluorescent spectrometry or potentiometry;
- (b) separating said tagged molecules by size;
- (c) cleaving said tag from said tagged molecule;
- and
- (d) detecting said tag by non-fluorescent spectrometry or potentiometry, and therefrom determining the identity of said nucleic acid molecule.
A method for detecting a selected nucleic acid molecule, comprising sequentially:
- (a) combining a tagged nucleic acid probe with target nucleic acid molecules under conditions and for a time sufficient to permit hybridization of said tagged nucleic acid probe to a complementary selected target nucleic acid sequence, wherein said tagged nucleic acid probe is detectable by non-fluorescent spectrometry or potentiometry;
- (b) increasing or decreasing the size of said hybridized tagged probes, the size of unhybridized probes or target molecules, or the size of the probe:target hybrids;
- (c) separating the tagged probes by size;
- (d) cleaving said tag from said tagged probe;
- and
- (e) detecting said tag by non-fluorescent spectrometry or potentiometry, and therefrom detecting said selected nucleic acid molecule.
A method for genotyping a selected organism, comprising sequentially:
- (a) generating tagged nucleic acid molecules from a selected target molecule, wherein a tag is correlative with a particular fragment and may be detected by non-fluorescent spectrometry or potentiometry;
- (b) separating said tagged molecules by sequential length;
- (c) cleaving said tag from said tagged molecule;
- and
- (d) detecting said tag by non-fluorescent spectrometry or potentiometry, and therefrom determining the genotype of said organism.
A method for genotyping a selected organism, comprising sequentially:
- (a) combining a tagged nucleic acid molecule with a selected target molecule under conditions and for a time sufficient to permit hybridization of said tagged molecule to said target molecule, wherein a tag is correlative with a particular fragment and may be detected by non-fluorescent spectrometry or potentiometry;
- (b) separating said tagged molecules by sequential length;
- (c) cleaving said tag from said tagged molecule;
- and
- (d) detecting said tag by non-fluorescent spectrometry or potentiometry, and therefrom determining the genotype of said organism.
A method for detecting a single selected nucleotide within a nucleic acid molecule, comprising sequentially:
- (a) hybridizing in at least two separate reactions a tagged primer and a target nucleic acid molecule under conditions and for a time sufficient to permit hybridization of said primer to said target nucleic acid molecule, wherein each reaction contains an enzyme which will add a nucleotide chain terminator, and, a nucleotide chain terminator complementary to adenosine, cytosine, guanosine, thymidine or uracil, and wherein each reaction contains a different nucleotide chain terminator;
- (b) separating said tagged primers by size;
- (c) cleaving said tag from said tagged primer;
- and
- (d) detecting said tag by non-fluorescent spectrometry or potentiometry, and therefrom determining the presence of said selected nucleotide within said nucleic acid molecule.
Dependencies are inferred from the claim text.