CA2386518A1 - Method and device for identifying a biopolymer sequence on a solid surface - Google Patents
Method and device for identifying a biopolymer sequence on a solid surface Download PDFInfo
- Publication number
- CA2386518A1 CA2386518A1 CA002386518A CA2386518A CA2386518A1 CA 2386518 A1 CA2386518 A1 CA 2386518A1 CA 002386518 A CA002386518 A CA 002386518A CA 2386518 A CA2386518 A CA 2386518A CA 2386518 A1 CA2386518 A1 CA 2386518A1
- Authority
- CA
- Canada
- Prior art keywords
- biopolymer
- change
- alternating
- affinity
- substrate
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6816—Hybridisation assays characterised by the detection means
- C12Q1/6825—Nucleic acid detection involving sensors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
- G01N33/542—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with steric inhibition or signal modification, e.g. fluorescent quenching
Abstract
The invention relates to a method for identifying a biopolymer spread on a first surface of a solid substrate, whereby the first biopolymer is brought into contact with a second biopolymer which spread on a second surface, whereby said second biopolymer has an affinity for the first biopolymer; Biopolymer in Kontakt gebracht wird und wobei die Identifikation des ersten Biopolymers vorgenommen wird durch Auswertung der durch affinitätsbedingte Adhäsion ausgelösten Anderung der Impedanz, der Leitfähigkeit im Gleichstrom - und/oder Wechselstrombereich in Abhängigkeit von einer aufgeprägten Wechselspannungs- oder Wechselstromfrequenz.
Description
Process and apparatus for the identification of a biopolysner sequence on the surfaces of solids The invention relates to a process and an apparatus for the identification of a specific biopolymer sequence which is bound to the surface of a solid.
US 5,780,234 discloses detecting the state of hybrid-ization by changing the electrical conductivity. To this end, it is necessary, according to the teaching of US 5,780,234, that transfer of free electrons takes place. For this purpose, the nucleic acid sequences are combined with electron donors or acceptors. If hybridization takes place, charge transport can occur.
The adduction of the oligonucleotide to be detected takes place from solution here.
Further processes for the identification of a polymer sequence are disclosed in WO 99/29898, US 5,065,798, WO 98/48275, US 5,866,336 and WO 99/11813.
It is known from H.W. Fink, C. Schonenberger, Nature 398, 407 (1999) that the conductivity of individual DNA
double strands is in the order of magnitude of good semiconductors or conductive polymers.
SØ Kelley, N.M. Jackson, M.G. Hill, J.K. Barton, Angew. Chem. Inter. Ed. Engl. 38, 941 (1999) disclose that monolayers of DNA double strands on electrode surfaces have high conductivity and faster charge transfer, even over large separations.
The object of the invention is to provide a novel technology by means of which biopolymers immobilized on a solid surface can be identified unambiguously, quickly and sensitively.
US 5,780,234 discloses detecting the state of hybrid-ization by changing the electrical conductivity. To this end, it is necessary, according to the teaching of US 5,780,234, that transfer of free electrons takes place. For this purpose, the nucleic acid sequences are combined with electron donors or acceptors. If hybridization takes place, charge transport can occur.
The adduction of the oligonucleotide to be detected takes place from solution here.
Further processes for the identification of a polymer sequence are disclosed in WO 99/29898, US 5,065,798, WO 98/48275, US 5,866,336 and WO 99/11813.
It is known from H.W. Fink, C. Schonenberger, Nature 398, 407 (1999) that the conductivity of individual DNA
double strands is in the order of magnitude of good semiconductors or conductive polymers.
SØ Kelley, N.M. Jackson, M.G. Hill, J.K. Barton, Angew. Chem. Inter. Ed. Engl. 38, 941 (1999) disclose that monolayers of DNA double strands on electrode surfaces have high conductivity and faster charge transfer, even over large separations.
The object of the invention is to provide a novel technology by means of which biopolymers immobilized on a solid surface can be identified unambiguously, quickly and sensitively.
- 2 This object is achieved by the features of Claims 1 and 7. Advantageous embodiments arise from the features of claims 2 to 6 and 8 to 12.
According to the invention, a process is provided for the identification of a first biopolymer applied to a first surface of a first substrate, where the first biopolymer is brought into contact with a second biopolymer which has affinity thereto and which is located on a surface of a second substrate, and where the identification of the first biopolymer is carried out by evaluation of the change in impedance or conductivity caused by the adhesion due to the affinity.
The term biopolymer is taken to mean, in particular, a polymer formed from nucleotides or amino acids, for example DNA, RNA, PNA, PTO, peptide, protein and the like. The term biopolymer which has affinity is taken to mean a biopolymer which is able to form a bond to a corresponding biopolymer. The bond can be a covalent, ionic or hydrogen bond. Bonding caused by steric effects is also possible.
Electronic detection of the change in conductivity between two surfaces which occurs, for example, on hybridization increases the sensitivity and specificity and reduces the apparatus complexity.
Possible areas of application of the process according to the invention are in medical diagnostics, identification, coding and recognition technology.
The change that can be measured is the change in impedance or conductivity in the direct-current and/or alternating-current region as a function of a superimposed alternating-voltage or current frequency.
It is possible here for one of the surfaces to be ~O 01/13115 PCT/DE00/02758
According to the invention, a process is provided for the identification of a first biopolymer applied to a first surface of a first substrate, where the first biopolymer is brought into contact with a second biopolymer which has affinity thereto and which is located on a surface of a second substrate, and where the identification of the first biopolymer is carried out by evaluation of the change in impedance or conductivity caused by the adhesion due to the affinity.
The term biopolymer is taken to mean, in particular, a polymer formed from nucleotides or amino acids, for example DNA, RNA, PNA, PTO, peptide, protein and the like. The term biopolymer which has affinity is taken to mean a biopolymer which is able to form a bond to a corresponding biopolymer. The bond can be a covalent, ionic or hydrogen bond. Bonding caused by steric effects is also possible.
Electronic detection of the change in conductivity between two surfaces which occurs, for example, on hybridization increases the sensitivity and specificity and reduces the apparatus complexity.
Possible areas of application of the process according to the invention are in medical diagnostics, identification, coding and recognition technology.
The change that can be measured is the change in impedance or conductivity in the direct-current and/or alternating-current region as a function of a superimposed alternating-voltage or current frequency.
It is possible here for one of the surfaces to be ~O 01/13115 PCT/DE00/02758
- 3 electrically conductive and for the change over this surface to be measured. However, the surfaces may also be separated by an insulator. The first and/or second biopolymer may be applied to the surface in the form of a layer, with electro-active metal atoms, ions, clusters or complex molecules being introduced therein.
According to a further embodiment, it is provided that the determination of the change is carried out by means of a reference electrode and/or a counterelectrode.
The invention furthermore relates to an apparatus for carrying out the process according to the invention, where a first biopolymer is applied to the surface of a first substrate in such a way that it can be brought into contact with a second biopolymer which has affinity thereto and which has been applied to a surface of a second substrate, and where, for the identification of the first biopolymer, a device is provided for the evaluation of a change in the impedance or conductivity caused by the adhesion due to the affinity.
Embodiments of the invention are explained below. The identification according to the invention of the biopolymer on the surface of a solid is advantageously carried out by the following procedure:
The first biopolymers immobilized on the surface of the first substrate are brought into contact with second biopolymers which have affinity thereto. This results in hybridization. The second biopolymers may be immobilized on a surface of a second substrate and be brought into contact with the first biopolymers to be identified, for example by pressing onto one another. A
suitable substrate material is glass, plastic or metal.
The latter may be in the form of a foil.
According to a further embodiment, it is provided that the determination of the change is carried out by means of a reference electrode and/or a counterelectrode.
The invention furthermore relates to an apparatus for carrying out the process according to the invention, where a first biopolymer is applied to the surface of a first substrate in such a way that it can be brought into contact with a second biopolymer which has affinity thereto and which has been applied to a surface of a second substrate, and where, for the identification of the first biopolymer, a device is provided for the evaluation of a change in the impedance or conductivity caused by the adhesion due to the affinity.
Embodiments of the invention are explained below. The identification according to the invention of the biopolymer on the surface of a solid is advantageously carried out by the following procedure:
The first biopolymers immobilized on the surface of the first substrate are brought into contact with second biopolymers which have affinity thereto. This results in hybridization. The second biopolymers may be immobilized on a surface of a second substrate and be brought into contact with the first biopolymers to be identified, for example by pressing onto one another. A
suitable substrate material is glass, plastic or metal.
The latter may be in the form of a foil.
- 4 When hybridization takes place between two parallel surfaces, the change in the conductivity between the two surfaces can be used for detection. This applies in both embodiments both for direct-current and alternating-current conductivity phenomena. In order to increase the conductivity, metal atoms, ions, clusters or complex molecules may also be intercalated into the thin films formed from the biopolymers. Alternatively, the detection can also take place via fluorescence or other optical methods. Conductive clusters can be employed here for amplification of optical signals.
In an embodiment, nucleic acids of a certain sequence are covalently bonded, as first biopolymers to be identified, to a conductive surface of the first substrate. Complementary nucleic acids are bound to a second conductive substrate, which is brought into contact with the first by pressing onto one another. If hybridization of the nucleic acids takes place, the electrical resistance drops. This can be detected by conventional electronic methods.
It is also possible to detect the changes in the capacitances in the hybridizing layer alternating-current resistances which accompany the hybridization.
Furthermore, it is also possible to employ the use of electrochemical signals, such as, for example, specific reduction and oxidation peaks, for identification of the hybridization.
The electronic measurement quantities can be amplified by introducing metal atoms, clusters or ions into the layer of the biopolymers to be detected. This can take place either before or after the hybridization, for example by vapor deposition or electrochemical methods.
Furthermore, it is also possible to use complex molecules which add specifically onto single-stranded structures or also as intercalators onto double-~n10 01/13115 PCT/DE00/02758
In an embodiment, nucleic acids of a certain sequence are covalently bonded, as first biopolymers to be identified, to a conductive surface of the first substrate. Complementary nucleic acids are bound to a second conductive substrate, which is brought into contact with the first by pressing onto one another. If hybridization of the nucleic acids takes place, the electrical resistance drops. This can be detected by conventional electronic methods.
It is also possible to detect the changes in the capacitances in the hybridizing layer alternating-current resistances which accompany the hybridization.
Furthermore, it is also possible to employ the use of electrochemical signals, such as, for example, specific reduction and oxidation peaks, for identification of the hybridization.
The electronic measurement quantities can be amplified by introducing metal atoms, clusters or ions into the layer of the biopolymers to be detected. This can take place either before or after the hybridization, for example by vapor deposition or electrochemical methods.
Furthermore, it is also possible to use complex molecules which add specifically onto single-stranded structures or also as intercalators onto double-~n10 01/13115 PCT/DE00/02758
- 5 stranded conformations and which have electro-active centers.
The process according to the invention can be used, for example, in the area of security technology for the marking and identification of bank notes, smart cards, identity cards and the like to prevent counterfeiting.
In the case of identification using a liquid phase, the process can also be employed for the marking and identification of, for example, foods, medicaments or the like.
Example:
Oligonucleotides having a length of 21 bases are covalently immobilized at the 5' -end to the surface of a conductive polycarbonate/carbon fiber plastic.
The oligonucleotides located on the surface are 20. hybridized with complementary probes located on a second surface. This is carried out by bringing the two surfaces into contact. If an alternating voltage having a frequency of 250 Hz is applied between the conductive plastic surface and the second surface and the capacitive proportion of the alternating current is measured, a drop in the alternating-current conductivity by more than 10~ arises in the case of hybridization. Hybridization of the oligonucleotide can thus be detected. Control experiments using non-specific oligonucleotides do not produce any significant change in conductivity.
The process according to the invention can be used, for example, in the area of security technology for the marking and identification of bank notes, smart cards, identity cards and the like to prevent counterfeiting.
In the case of identification using a liquid phase, the process can also be employed for the marking and identification of, for example, foods, medicaments or the like.
Example:
Oligonucleotides having a length of 21 bases are covalently immobilized at the 5' -end to the surface of a conductive polycarbonate/carbon fiber plastic.
The oligonucleotides located on the surface are 20. hybridized with complementary probes located on a second surface. This is carried out by bringing the two surfaces into contact. If an alternating voltage having a frequency of 250 Hz is applied between the conductive plastic surface and the second surface and the capacitive proportion of the alternating current is measured, a drop in the alternating-current conductivity by more than 10~ arises in the case of hybridization. Hybridization of the oligonucleotide can thus be detected. Control experiments using non-specific oligonucleotides do not produce any significant change in conductivity.
Claims (10)
1. A process for the identification of an anti-counterfeiting mark in the form of a first biopolymer applied to a surface of a first substrate, where the first biopolymer is applied to the surface as a layer with electro-active metal atoms, ions, clusters or complex molecules introduced therein in order to amplify electrical measurement quantities, where the first biopolymer is brought into contact with a second biopolymer which has affinity thereto and which has been applied to a surface of a second substrate as a layer with electro-active metal atoms, ions, clusters or complex molecules introduced therein in order to amplify electrical measurement quantities, and where the identification of the first biopolymer is carried out by evaluation of the change in the electrical measurement quantity impedance or conductivity caused by the adhesion due to the affinity.
2. A process as claimed in claim 1, where the change in the impedance or conductivity in the direct- and/or alternating-current region is measured as a function of a superimposed alternating-voltage or alternating-current frequency.
3. A process as claimed in claim 1 or 2, where at least one of the surfaces is electrically conductive, and the change over this surface is measured.
4. A process as claimed in claim 3, where the surfaces are separated by an insulator.
5. A process as claimed in the preceding claims, where the determination of the change is carried out by means of a reference electrode and/or a counter-electrode.
6. An apparatus for carrying out the process as claimed in one of the preceding claims, where a first biopolymer is applied to a surface of a first substrate in such a way that it can be brought into contact with a second biopolymer which has affinity thereto and which has been applied to a surface of a second substrate, where the first and second biopolymer are on the surface in the form of a layer, and the layer is provided with electro-active metal atoms, ions, clusters or complex molecules in order to amplify electrical measurement quantities, and where, for the identification of the first biopolymer, a device is provided for the evaluation of a change in the electrical measurement quantity impedance or conductivity caused by the adhesion due to the affinity.
7. An apparatus as claimed in claim 6, where the change in the direct-current and/or alternating-current region as a function of a superimposed alternating-voltage or alternating-current frequency can be measured by means of the evaluation device.
8. An apparatus as claimed in claim 6 or 7, where at least one of the surfaces is electrically conductive, and the change can be measured over this surface.
9. An apparatus as claimed in one of claims 6 to 8, where the first and/or second biopolymer has been applied to an insulator provided on the surface.
10. An apparatus as claimed in one of claims 6 to 9, where a reference electrode and/or a counterelectrode is/are provided.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19938138.0 | 1999-08-16 | ||
| DE19938138A DE19938138C2 (en) | 1999-08-16 | 1999-08-16 | Method and device for identifying a biopolymer sequence on solid surfaces |
| PCT/DE2000/002758 WO2001013115A2 (en) | 1999-08-16 | 2000-08-12 | Method and device for identifying a biopolymer sequence on a solid surface |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2386518A1 true CA2386518A1 (en) | 2001-02-22 |
Family
ID=7918107
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002386518A Abandoned CA2386518A1 (en) | 1999-08-16 | 2000-08-12 | Method and device for identifying a biopolymer sequence on a solid surface |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP1203237A2 (en) |
| AU (1) | AU7403000A (en) |
| CA (1) | CA2386518A1 (en) |
| DE (1) | DE19938138C2 (en) |
| WO (1) | WO2001013115A2 (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE10113711A1 (en) * | 2001-03-16 | 2002-09-26 | Lifebits Ag | Identifying bio-chemical reactions e.g., DNA hybridization, comprises digitally scanning specific detector molecules in an array of accurately-defined and discrete points with a sample, for statistical evaluation |
| DE10113550A1 (en) * | 2001-03-20 | 2002-10-02 | Infineon Technologies Ag | Method for detecting macromolecular biopolymers using an electrode arrangement |
| US8148164B2 (en) | 2003-06-20 | 2012-04-03 | Roche Diagnostics Operations, Inc. | System and method for determining the concentration of an analyte in a sample fluid |
| US7645421B2 (en) | 2003-06-20 | 2010-01-12 | Roche Diagnostics Operations, Inc. | System and method for coding information on a biosensor test strip |
| US7645373B2 (en) | 2003-06-20 | 2010-01-12 | Roche Diagnostic Operations, Inc. | System and method for coding information on a biosensor test strip |
| US7718439B2 (en) | 2003-06-20 | 2010-05-18 | Roche Diagnostics Operations, Inc. | System and method for coding information on a biosensor test strip |
| US8206565B2 (en) | 2003-06-20 | 2012-06-26 | Roche Diagnostics Operation, Inc. | System and method for coding information on a biosensor test strip |
| US8058077B2 (en) | 2003-06-20 | 2011-11-15 | Roche Diagnostics Operations, Inc. | Method for coding information on a biosensor test strip |
| US7569126B2 (en) | 2004-06-18 | 2009-08-04 | Roche Diagnostics Operations, Inc. | System and method for quality assurance of a biosensor test strip |
Family Cites Families (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2598227B1 (en) * | 1986-04-30 | 1989-07-28 | Bio Merieux | METHOD FOR DETECTION AND / OR IDENTIFICATION OF A BIOLOGICAL SUBSTANCE IN A LIQUID MEDIUM USING ELECTRICAL MEASUREMENTS, AND DEVICE FOR CARRYING OUT SAID METHOD |
| US5605798A (en) * | 1993-01-07 | 1997-02-25 | Sequenom, Inc. | DNA diagnostic based on mass spectrometry |
| US5591578A (en) * | 1993-12-10 | 1997-01-07 | California Institute Of Technology | Nucleic acid mediated electron transfer |
| US5567301A (en) * | 1995-03-01 | 1996-10-22 | Illinois Institute Of Technology | Antibody covalently bound film immunobiosensor |
| US5866336A (en) * | 1996-07-16 | 1999-02-02 | Oncor, Inc. | Nucleic acid amplification oligonucleotides with molecular energy transfer labels and methods based thereon |
| GB9622304D0 (en) * | 1996-10-26 | 1996-12-18 | Univ Manchester | Sensor |
| US6306584B1 (en) * | 1997-01-21 | 2001-10-23 | President And Fellows Of Harvard College | Electronic-property probing of biological molecules at surfaces |
| US6669906B1 (en) * | 1997-04-22 | 2003-12-30 | Thomas Schalkhammer | Reinforced cluster optical sensors |
| EP1009852A2 (en) * | 1997-09-04 | 2000-06-21 | Bayer Corporation | Oligonucleotide probes bearing quenchable fluorescent labels, and methods of use thereof |
| WO1999029898A2 (en) * | 1997-12-05 | 1999-06-17 | MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. | Method for identifying nucleic acids by means of matrix-assisted laser desorption/ionisation mass spectrometry |
| JP2003508730A (en) * | 1999-06-10 | 2003-03-04 | モトローラ・インコーポレイテッド | Biosensor using charge neutral conjugated polymer |
-
1999
- 1999-08-16 DE DE19938138A patent/DE19938138C2/en not_active Expired - Fee Related
-
2000
- 2000-08-12 AU AU74030/00A patent/AU7403000A/en not_active Abandoned
- 2000-08-12 WO PCT/DE2000/002758 patent/WO2001013115A2/en not_active Application Discontinuation
- 2000-08-12 EP EP00962216A patent/EP1203237A2/en not_active Withdrawn
- 2000-08-12 CA CA002386518A patent/CA2386518A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| DE19938138C2 (en) | 2003-02-13 |
| WO2001013115A3 (en) | 2001-08-23 |
| DE19938138A1 (en) | 2001-03-01 |
| AU7403000A (en) | 2001-03-13 |
| WO2001013115A2 (en) | 2001-02-22 |
| EP1203237A2 (en) | 2002-05-08 |
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Legal Events
| Date | Code | Title | Description |
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| FZDE | Dead |