Claims
- 1. A universal gel comprising a universal capture probe comprising a nucleotide sequence region complementary to a capture probe-specific nucleotide sequence region contained within an adapter molecule, wherein said capture probe is covalently immobilized within a medium suitable for electrophoresis.
- 2. The universal gel of claim 1, wherein said universal capture probe comprises SEQ ID No. 5.
- 3. The universal gel of claim 2, wherein an adapter molecule is selected from the group consisting of: SEQ ID NOS. 1 and 2.
- 4. A universal capture gel system comprising:
(a) a universal capture probe, wherein said universal capture probe comprises a nucleotide sequence region complementary to said adapter molecule; (b) an adapter molecule, wherein said adapter molecule comprises a capture probe-specific nucleotide sequence region which is complementary to a nucleotide sequence region contained within said universal capture probe, and a target-specific nucleotide sequence region complementary to a nucleotide sequence region contained within a target molecule; and (c) a medium suitable for electrophoresis, wherein said universal capture probe is covalently immobilized within said medium suitable for electrophoresis.
- 5. A universal gel hybridization complex comprising:
(a) at least one adapter molecule comprising a capture probe-specific nucleotide sequence region that is complementary to a nucleotide sequence region contained within a universal capture probe, and a target-specific nucleotide sequence region that is complementary to a nucleotide sequence region of at least one target molecule; and (b) at least one target molecule comprising a nucleotide sequence complementary to said adapter's target-specific nucleotide sequence region of (a), wherein the adapter molecule of (a) is hybridized with the target molecule of (b) thereby forming a hybridization complex.
- 6. The method of claim 5, wherein said universal capture probe is SEQ ID No. 5.
- 7. The universal gel hybridization complex of claim 5, wherein said adapter molecule contains from about 10 to about 100 nucleotides.
- 8. The universal gel hybridization complex of claim 5, wherein said adapter molecule is either a single-stranded or double-stranded nucleic acid.
- 9. The adapter molecule of claim 8, wherein said nucleic acid is either DNA or RNA.
- 10. The universal gel hybridization complex of claim 5, wherein said adapter molecule is an aptamer.
- 11. The universal gel hybridization complex of claim 5, wherein said target molecule is selected from the group consisting of: nucleic acids, nucleic acid analogs, modified nucleic acids, aptamer binding partners and nucleic acid binding proteins.
- 12. The universal gel hybridization complex of claim 5, wherein said target molecule is either a single-stranded or double-stranded nucleic acid, modified nucleic acid, or nucleic acid analog molecule.
- 13. The universal gel hybridization complex of claim 5, wherein said target molecule is ribonucleic acid.
- 14. The universal gel hybridization complex of claim 5, wherein said target molecule is deoxyribonucleic acid.
- 15. The universal gel hybridization complex of claim 5, wherein said target molecule contains from about 10 to about 100,000 nucleotides.
- 16. The universal gel hybridization complex of claim 5, wherein the test sample comprises a target molecule selected from the group consisting of: bacterial molecules, viral molecules, fungal molecules, parasitic molecules, plant molecules, animal molecules and combinations thereof.
- 17. A method of detecting the presence, or absence, of a target molecule in a test sample, comprising the following steps:
(a) forming a hybridization complex by contacting:
(i) an adapter molecule comprising a capture probe-specific nucleotide sequence region that is complementary to a nucleotide sequence region contained within at least one universal capture probe polynucleotide immobilized within an electrophoresis medium, and a target-specific nucleotide sequence region that is complementary to a nucleotide sequence region contained within at least one target molecule; and (ii) the test sample wherein the test sample contains a target molecule comprising a nucleotide sequence complementary to said adapter's target-specific nucleotide sequence region, under conditions suitable for hybridization of said adapter molecule with said target molecule; (b) introducing said hybridization complex of (a) into an electrophoresis medium comprising an immobilized universal capture probe wherein the capture probe comprises a nucleotide sequence region complementary to said adapter's capture probe-specific nucleotide sequence region; (c) subjecting said electrophoresis medium to an electric field resulting in the electrophoretic migration of the hybridization complex formed in (a) into at least one region of said medium containing at least one class of immobilized universal capture probes, thereby forming an adapter/target/universal capture probe complex; and (d) detecting said adapter/target/universal capture probe complex, wherein the detection of said adapter/target/universal capture probe complex is indicative of the presence of at least one target molecule within said test sample.
- 18. The method of claim 17, wherein the electrophoretic medium used in said universal gel is selected from the group consisting of: a polyacrylamide polymer, an agarose polymer, a starch polymer and a combination thereof.
- 19. The method of claim 17, wherein said target molecule is labeled.
- 20. The method of claim 17, wherein said label is selected from the group consisting of: radioactivity, catalytic, chemiluminescent, phosphorescent, fluorescent and luminescent.
- 21. The method of claim 17, wherein said universal capture probes are immobilized within at least one discrete region of the electrophoresis medium.
- 22. The method of claim 17, wherein said universal capture probes are immobilized throughout said electrophoresis medium.
- 23. The method of claim 17, wherein said universal capture probe is selected from the group consisting of: nucleic acids, nucleic acid analogs, and modified nucleic acids.
- 24. The method of claim 17, wherein said universal capture probe is SEQ ID NO. 5.
- 25. The method of claim 17, wherein said universal capture probe contains from about 5 to about 100 nucleotides.
- 26. The method of claim 17, wherein said target molecule is selected from the group consisting of: nucleic acids, nucleic acid analogs, modified nucleic acids, aptamer binding partners and nucleic acid binding proteins.
- 27. The method of claim 17, wherein said target molecule is either a single-stranded or double-stranded nucleic acid, modified nucleic acid, or nucleic acid analog molecule.
- 28. The method of claim 17, wherein said target molecule is ribonucleic acid.
- 29. The method of claim 17, wherein said target molecule is deoxyribonucleic acid.
- 30. The method of claim 17, wherein said target molecule contains from about 10 to about 100,00 nucleotides.
- 31. The method of claim 17, wherein the test sample comprises a target molecule selected from the group consisting of: bacterial molecules, viral molecules, fungal molecules, parasitic molecules, plant molecules, animal molecules and combinations thereof.
- 32. A method of detecting the presence or absence of a target molecule in a test sample, comprising the following steps:
(a) introducing said test sample into an electrophoretic medium comprising immobilized capture probes; (b) subjecting said electrophoretic medium of (a) to an electric field resulting in the electrophoretic migration of target molecules of the test sample into said electrophoretic medium; (c) introducing an adapter molecule into said electrophoretic medium of (a), wherein said adapter is subjected to electrophoretic migration through said electrophoretic medium, wherein said adapter molecule comprises an electrophoretic mobility greater than said target molecule; (d) contacting said adapter with said target molecule within said electrophoretic medium, thereby forming an adapter/target complex; (e) contacting said adapter/target complex of (d), with said immobilized capture probe, thereby forming an adapter/target/capture probe complex; and (f) detecting said adapter/target/universal capture probe complex of (e), wherein the detection of said adapter/target/universal capture probe complex is indicative of the presence of at least one target molecule within said test sample.
- 33. A method of detecting the presence or absence of a target molecule in a test sample, comprising the following steps:
(a) introducing an adapter molecule into an electrophoretic medium comprising immobilized capture probes; (b) subjecting said electrophoretic medium to an electric field resulting in the electrophoretic migration of said adapter molecule into said medium comprising immobilized capture probes, thereby forming an adapter/capture probe complex; (c) introducing said test sample into said electrophoretic medium of (b), wherein target molecules of the test sample are subjected to electrophoretic migration into said electrophoretic medium, thereby forming an adapter/target/capture probe complex; and (d) detecting said adapter/target/capture probe complex of (c), wherein the detection of said adapter/target/capture probe complex is indicative of the presence of at least one target molecule within said test sample.
- 34. A method of detecting the presence, or absence, of one, or more, target molecules in a test sample, comprising the following steps:
(a) forming multiple hybridization complexes by contacting:
(i) appropriate adapter molecules, wherein each adapter molecule comprises a capture probe-specific nucleotide sequence region that is complementary to a nucleotide sequence region contained within a universal capture probe immobilized within an electrophoresis medium, and a target-specific nucleotide sequence region that is complementary to a nucleotide sequence region contained within a specific target molecule; and (ii) one or more target molecules, wherein each target molecule comprises a nucleotide sequence complementary to an adapter's target-specific nucleotide sequence region, under conditions suitable for hybridization of said adapter molecule said target molecule; (b) introducing said hybridization complexes of (a) into an electrophoretic medium comprising universal capture probes immobilized within one, or more, discrete regions of said medium; (c) subjecting said electrophoretic medium to an electric field resulting in the electrophoretic migration of the hybridization complexes formed in (a) into at least one region of said universal gel containing only one class of universal capture probes immobilized in one, or more, discrete regions of said universal gel, thereby forming adapter/target/universal capture probe complexes; and (d) detecting said adapter/target/universal capture probe complex, wherein the detection of said adapter/target/universal capture probe complex is indicative of the presence of at least one target molecule within said test sample.
- 35. A method of purifying a target molecule from a test sample, comprising the following steps:
(a) forming a hybridization complex by contacting:
(i) an adapter molecule comprising a capture probe-specific nucleotide sequence region that is complementary to a nucleotide sequence region contained within at least one universal capture probe polynucleotide immobilized within an electrophoresis medium, and a target-specific nucleotide sequence region that is complementary to a nucleotide sequence region contained within at least one target molecule; and (ii) the test sample wherein the test sample contains a target molecule comprising a nucleotide sequence complementary to said adapter's target-specific nucleotide sequence region, under conditions suitable for hybridization of said adapter molecule with said target molecule; (b) introducing said hybridization complex of (a) into an electrophoresis medium comprising an immobilized universal capture probe wherein the capture probe comprises a nucleotide sequence region complementary to said adapter's capture probe-specific nucleotide sequence region; (c) subjecting said electrophoresis medium to an electric field resulting in the electrophoretic migration of the hybridization complex formed in (a) into at least one region of said medium containing at least one class of immobilized universal capture probes, thereby forming an adapter/target/universal capture probe complex; (d) introducing a modified adapter molecule to said electrophoresis medium, wherein said modified adapter molecule comprises a nucleotide sequence region complementary to the unmodified adapter's target-specific nucleotide region; (e) subjecting said modified adapter molecule to electrophoresis by applying an electric field to said electrophoresis medium; and (f) displacing said target molecule from said adapter/target/universal capture probe complex by contacting said unmodified adapter with said adapter/target/universal capture probe complex, wherein said modified adapter displaces said target and hybridizes to said adapter molecule.
- 36. The method of claim 35 wherein in step (d), said modified adapter comprises a nucleotide sequence region complementary to said unmodified adapter's capture probe-specific nucleotide sequence, and wherein in step (f), the target/adapter complex is displaced from the adapter/target/universal capture probe complex by contacting said modified adapter, under conditions suitable for hybridization.
RELATED APPLICATIONS
[0001] This application is a continuation of PCT/US00/08529, filed Mar. 31, 2000, which claims priority to U.S. application Ser. No. 09/285,380, filed Apr. 2, 1999, which is a continuation in part of U.S. application Ser. No. 08/971,845, filed Aug. 8, 1997, which claims the benefit of Provisional Application No. 60/046,708, filed May 16, 1997, the entire teachings of which are incorporated herein by reference.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60046708 |
May 1997 |
US |
Continuations (2)
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Number |
Date |
Country |
Parent |
PCT/US00/08529 |
Mar 2000 |
US |
Child |
09968084 |
Oct 2001 |
US |
Parent |
09285380 |
Apr 1999 |
US |
Child |
PCT/US00/08529 |
Mar 2000 |
US |
Continuation in Parts (1)
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Number |
Date |
Country |
Parent |
08971845 |
Aug 1997 |
US |
Child |
09285380 |
Apr 1999 |
US |