WO1998048048A2 - Dna mutation mapping by multiple energy transfer interactions - Google Patents

Dna mutation mapping by multiple energy transfer interactions Download PDF

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Publication number
WO1998048048A2
WO1998048048A2 PCT/GB1998/001138 GB9801138W WO9848048A2 WO 1998048048 A2 WO1998048048 A2 WO 1998048048A2 GB 9801138 W GB9801138 W GB 9801138W WO 9848048 A2 WO9848048 A2 WO 9848048A2
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markers
seq
target sequence
oligonucleotides
sequence
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PCT/GB1998/001138
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French (fr)
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WO1998048048A3 (en
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Shankar Balasubramanian
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Cambridge University Technical Services Ltd.
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Priority to AU70641/98A priority Critical patent/AU7064198A/en
Publication of WO1998048048A2 publication Critical patent/WO1998048048A2/en
Publication of WO1998048048A3 publication Critical patent/WO1998048048A3/en

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    • 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
    • C12Q1/6818—Hybridisation assays characterised by the detection means involving interaction of two or more labels, e.g. resonant energy transfer

Definitions

  • This invention relates to DNA mismatch screening, especially using FRET-detected hybridisation.
  • One of the simplest methods for detecting gene sequences is to make use of the specific hybridisation reaction between the target sequence and a suitable probe.
  • FRET Fluorescence Transfer
  • the probes each comprise a stem-and-loop structure with the stem formed by the annealing of two complementary arm sequences either side of the probe sequence.
  • a fluorescent moiety is attached to one arm and a non-fluorescent quenching moiety is attached to the other arm. Separation of the stem structure occurs on hybridisation of the probe to the complementary target sequence. This separates the two moieties and allows fluorescence to occur.
  • a method for determining the presence of a mismatch in a target sequence comprises contacting the target sequence with first, second and third oligonucleotides capable of hybridising to the natural sequence, in juxtaposition, wherein the oligonucleotides are respectively labelled with first, second and third markers having first, second and third absorption wavelengths and first, second and third emission wavelengths such that there is resonance between either or each of first and second markers and between second and third markers, and observing the presence or absence of each or either resonance.
  • Fig. 1 is a schematic representation of the interaction of labelled oligonucleotides and target sequence in a screening method embodying the invention
  • Fig. 2 is a schematic representation of a more specific embodiment of the invention and represents another arrangement of the labels.
  • Fig. 3 illustrates the different emission intensities obtained when point mutations are present in the target sequence, using the arrangement of fluorophores shown in Fig. 2. Description of Invention
  • the ability to monitor the interactions between nucleic acids is achieved through the detection of sensitised acceptor emission, due to FRET between a donor fluorophore and two different acceptor fluorophores on associating strands.
  • sensitised acceptor emission due to FRET between a donor fluorophore and two different acceptor fluorophores on associating strands.
  • the absence of any of the components induced by a single base mismatch should cause a detectable loss in sensitised acceptor emission.
  • the system illustrated in Fig. 1 comprises a target strand and three adjacent complementary oligonucleotides, A, B and C. These are typically each 8-13 base pairs in length.
  • a and B are modified at the 5' end with a fluorescein moiety (F) .
  • B and C are modified at the 3' end with two distinct types of acceptor fluorophore (X and Y) having different emission maxima and absorption profiles which overlap with the emission profile of fluorescein.
  • Fig. 2 illustrates a different arrangement of the fluorophores.
  • three oligonucleotides are used.
  • the central oligonucleotide is labelled with a donor fluorophore, 5'-carboxyfluorescein (fluorescein) only.
  • the two adjacent oligonucleotides are each labelled with an individual acceptor fluorophore, 5'-carboxytetramethyl- rhodamine (TMR) and 5 ' -carboxyrhodamine-X (ROX) .
  • TMR 5'-carboxytetramethyl- rhodamine
  • ROX 5 ' -carboxyrhodamine-X
  • the fluorophores are positioned such that excitation of fluorescein results in energy transfer from fluorescein to both TMR and ROX. This transfer can again be monitored by observing the emission of the two acceptor fluorophores. Since the acceptors emit at two distinct wavelengths, introduction of a single base mismatch should be detectable by a loss in either or both of these signals.
  • the positioning of the fluorophores on each oligonucleotide required to optimise energy transfer can be easily determined by the skilled person by preliminary studies.
  • the fluorophores will typically be separated, e.g. by a distance of 7 bases.
  • FRET assays can be carried out using a fluorimager, irradiating the assays with a laser at a suitable wavelength, e.g. 488 nm for fluorescein, and scanning the emissions using suitable filters, e.g. 530 nm for fluorescein, 570 nm for TMR and 610 nm for ROX.
  • a fluorimager irradiating the assays with a laser at a suitable wavelength, e.g. 488 nm for fluorescein, and scanning the emissions using suitable filters, e.g. 530 nm for fluorescein, 570 nm for TMR and 610 nm for ROX.
  • a characteristic of the common, donor fluorophore is that it should emit at a wavelength that is capable of exciting each of the acceptor fluorophores. The emission of each acceptor fluorophore must be resolvable.
  • a common fluorophore that can be used as a donor is carboxyfluorescein.
  • suitable acceptors include N,N,N' ,N' -tetramethyl-6- carboxyrhodamine and 2 ' , 1 ' -dimethoxy-4 ' , 5 ' -dichloro-6- carboxyfluorescein.
  • the probes may be of DNA.
  • fluorescently-labelled DNA mimics e.g. PNA, phosphorothioate DNA, could be used as the probes.
  • Example 1 illustrates the invention. In particular, it shows the ability to detect and locate point mutations in a target oligonucleotide.
  • SEQ ID Nos 1-4 Four target oligonucleotides (SEQ ID Nos 1-4) were designed, each differing only in one nucleotide.
  • oligo SEQ ID No 1 As the control, three oligonucleotide probes (SEQ ID No. 5-7) were designed, each capable of hybridising to a distinct region on the control.
  • the probes were labelled with a fluorophore at the positions marked (*) as follows: probe SEQ ID No 5 with ROX; probe SEQ ID No 6 with fluorescein; and probe SEQ ID No 7 with TMR.
  • the emission intensity for both ROX and TMR-labelled probes increased significantly (relative to each singly- bound probe) on hybridisation to the control.
  • the emission intensity varied significantly, depending on the position of the single nucleotide difference from the control.
  • This assay provides a simple method for the analysis of hybridisation events in solution and enables detection of mutations within defined regions of a target strand from a single fluorescence measurement.
  • the assay may be modified to utilise immobilised target or probes.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
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  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Immunology (AREA)
  • Physics & Mathematics (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
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  • Analytical Chemistry (AREA)
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  • Bioinformatics & Cheminformatics (AREA)
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Abstract

A method for determining the presence and location of a mismatch in a target sequence using Fluorescence Resonance Energy Transfer (FRET). The method comprises contacting the target sequence with at least three labelled oligonucleotide probes capable of hybridising to the natural sequence, in juxtaposition. The central probe is labelled with a donor fluorophore and the probe either side of this is labelled with a distinct acceptor fluorophore. Hybridisation to the target sequence results in resonance between the donor fluorophore and the acceptor fluorophores. A mismatch present in the target sequence will disrupt hybridisation to that region, resulting in a significant alteration to the resonance signal.

Description

DNA MUTATION MAPPING BY MULTIPLE ENERGY TRANSFER INTERACTIONS Field of Invention
This invention relates to DNA mismatch screening, especially using FRET-detected hybridisation. Background of the Invention
There is a general increase in the exploration of gene sequences and function. This has generated a need for new approaches to nucleic acid analysis. One of the simplest methods for detecting gene sequences is to make use of the specific hybridisation reaction between the target sequence and a suitable probe.
Recently, the use of Fluorescence Resonance Energy
Transfer (FRET) has been applied to the detection of hybridised probes. Tyagi et al . , Nature Biotech. (1996)
14:303-308, describe the use of FRET to distinguish between hybridised and unhybridised probes in a homogeneous assay.
The probes each comprise a stem-and-loop structure with the stem formed by the annealing of two complementary arm sequences either side of the probe sequence. A fluorescent moiety is attached to one arm and a non-fluorescent quenching moiety is attached to the other arm. Separation of the stem structure occurs on hybridisation of the probe to the complementary target sequence. This separates the two moieties and allows fluorescence to occur. Summary of Invention
This invention is based on a realisation of the utility of FRET as a technique for mapping point mutations on a single strand of DNA. According to the present invention, a method for determining the presence of a mismatch in a target sequence, comprises contacting the target sequence with first, second and third oligonucleotides capable of hybridising to the natural sequence, in juxtaposition, wherein the oligonucleotides are respectively labelled with first, second and third markers having first, second and third absorption wavelengths and first, second and third emission wavelengths such that there is resonance between either or each of first and second markers and between second and third markers, and observing the presence or absence of each or either resonance. Description of the Drawings
The accompanying drawings are for the purpose of illustration only. In the drawings:
Fig. 1 is a schematic representation of the interaction of labelled oligonucleotides and target sequence in a screening method embodying the invention;
Fig. 2 is a schematic representation of a more specific embodiment of the invention and represents another arrangement of the labels; and
Fig. 3 illustrates the different emission intensities obtained when point mutations are present in the target sequence, using the arrangement of fluorophores shown in Fig. 2. Description of Invention
The ability to monitor the interactions between nucleic acids is achieved through the detection of sensitised acceptor emission, due to FRET between a donor fluorophore and two different acceptor fluorophores on associating strands. The absence of any of the components induced by a single base mismatch should cause a detectable loss in sensitised acceptor emission.
The system illustrated in Fig. 1 comprises a target strand and three adjacent complementary oligonucleotides, A, B and C. These are typically each 8-13 base pairs in length. A and B are modified at the 5' end with a fluorescein moiety (F) . B and C are modified at the 3' end with two distinct types of acceptor fluorophore (X and Y) having different emission maxima and absorption profiles which overlap with the emission profile of fluorescein.
When no mismatches are present, all three oligonucleotides will be bound to the target strand. Thus, when the system is excited at the absorption wavelength of fluorescein (488 nm) , FRET will occur and the fluorescence spectrum will show a decrease in the fluorescein emission relative to each singly-bound probe and appearance of two maxima due to secondary emissions from X and Y. In cases where a mismatch exists somewhere within the target sequence, the region in which the error occurs can be distinguished simply by analysing the fluorescence spectrum. If the mismatch lies within the binding domain of oligonucleotide A, no duplex will form in this region and the secondary emission from X will be lost. Similarly, for mismatches within the B region, the signals from X and Y will disappear, and for errors in region C, the Y emission will be absent.
Fig. 2 illustrates a different arrangement of the fluorophores. Again, three oligonucleotides are used. The central oligonucleotide is labelled with a donor fluorophore, 5'-carboxyfluorescein (fluorescein) only. The two adjacent oligonucleotides are each labelled with an individual acceptor fluorophore, 5'-carboxytetramethyl- rhodamine (TMR) and 5 ' -carboxyrhodamine-X (ROX) . These fluorophores have absorption spectra which overlap with fluorescein' s emission spectrum and have well separated emission maxima.
On binding of all three oligonucleotides, the fluorophores are positioned such that excitation of fluorescein results in energy transfer from fluorescein to both TMR and ROX. This transfer can again be monitored by observing the emission of the two acceptor fluorophores. Since the acceptors emit at two distinct wavelengths, introduction of a single base mismatch should be detectable by a loss in either or both of these signals.
The positioning of the fluorophores on each oligonucleotide required to optimise energy transfer can be easily determined by the skilled person by preliminary studies. The fluorophores will typically be separated, e.g. by a distance of 7 bases.
FRET assays can be carried out using a fluorimager, irradiating the assays with a laser at a suitable wavelength, e.g. 488 nm for fluorescein, and scanning the emissions using suitable filters, e.g. 530 nm for fluorescein, 570 nm for TMR and 610 nm for ROX.
A characteristic of the common, donor fluorophore is that it should emit at a wavelength that is capable of exciting each of the acceptor fluorophores. The emission of each acceptor fluorophore must be resolvable.
An example of a common fluorophore that can be used as a donor is carboxyfluorescein. Additional examples of suitable acceptors include N,N,N' ,N' -tetramethyl-6- carboxyrhodamine and 2 ' , 1 ' -dimethoxy-4 ' , 5 ' -dichloro-6- carboxyfluorescein.
The probes may be of DNA. Alternatively, and on the same principle, fluorescently-labelled DNA mimics, e.g. PNA, phosphorothioate DNA, could be used as the probes.
By the use of additional fluorophores with distinct emission profiles, it may be possible to extend this system for the screening of longer target sequences using multiple short complementary strands. This arrangement may permit screening of a section of the DNA target gene in a single homogeneous assay by excitation with, e.g. a fixed wavelength laser, or a fluorescence spectrometer, and instantly generating an emission spectrum which characterises where, if anywhere, there is a point mutation or damage. It is envisaged that sequences of diagnostic oligonucleotides may be chosen to generate diagnostic kits for screening regions of mutational "hotspots" .
The following Example illustrates the invention. In particular, it shows the ability to detect and locate point mutations in a target oligonucleotide. Example
Four target oligonucleotides (SEQ ID Nos 1-4) were designed, each differing only in one nucleotide.
SEQ ID No 1 5'CGTTCTAAGGATTACGTCGAACCTTTG3' SEQ ID No 2 5 ' CGTTCAAAGGATTACGTCGAACCTTTG3 '
SEQ ID No 3 5 ' CGTTCTAAGGATAACGTCGAACCTTTG3 ' SEQ ID No 4 5'CGTTCTAAGGATTACGTCGAACCATTG3' s
Using oligo SEQ ID No 1 as the control, three oligonucleotide probes (SEQ ID No. 5-7) were designed, each capable of hybridising to a distinct region on the control.
SEQ ID No 5 3 ' GCAAGATTC5 ' SEQ ID No 6 3 ' CTAATGCAG5 ' SEQ ID No 7 3 ' CTTGGAAAC5 '
The probes were labelled with a fluorophore at the positions marked (*) as follows: probe SEQ ID No 5 with ROX; probe SEQ ID No 6 with fluorescein; and probe SEQ ID No 7 with TMR. The emission intensity for both ROX and TMR-labelled probes increased significantly (relative to each singly- bound probe) on hybridisation to the control. However, as shown in Fig. 3, on reaction with each of the target oligonucleotides SEQ ID Nos 2-4, the emission intensity varied significantly, depending on the position of the single nucleotide difference from the control.
This assay provides a simple method for the analysis of hybridisation events in solution and enables detection of mutations within defined regions of a target strand from a single fluorescence measurement. The assay may be modified to utilise immobilised target or probes.
6
SEQUENCE LISTING
(1) GENERAL INFORMATION:
(i) APPLICANT:
(A) NAME: Cambridge University Technical Services Ltd.
(B) STREET: The Old Schools, Trinity Lane
(C) CITY: Cambridge
(D) STATE: N/A
(E) COUNTRY: United Kingdom
(F) POSTAL CODE (ZIP): CB2 ITS
(ii) TITLE OF INVENTION: DNA MUTATION MAPPING BY MULTIPLE ENERGY TRANSFER INTERACTIONS
(iii) NUMBER OF SEQUENCES: 7
(iv) COMPUTER READABLE FORM:
(A) MEDIUM TYPE: Floppy disk
(B) COMPUTER: IBM PC compatible
(C) OPERATING SYSTEM: PC-DOS/MS-DOS
(D) SOFTWARE: Patentln Release #1.0, Version #1.30 (EPO)
(v) CURRENT APPLICATION DATA:
APPLICATION NUMBER: WO Not yet known
(2) INFORMATION FOR SEQ ID NO: 1:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 27 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS : single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: other nucleic acid
(A) DESCRIPTION: /desc = "Oligonucleotide"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1: CGTTCTAAGG ATTACGTCGA ACCTTTG 27
(2) INFORMATION FOR SEQ ID NO: 2:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 27 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: other nucleic acid
(A) DESCRIPTION: /desc = "Oligonucleotide"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2: CGTTCAAAGG ATTACGTCGA ACCTTTG 27 s
(2) INFORMATION FOR SEQ ID NO: 3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 27 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: other nucleic acid
(A) DESCRIPTION: /desc = "Oligonucleotide"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3: CGTTCTAAGG ATAACGTCGA ACCATTG 27
(2) INFORMATION FOR SEQ ID NO: 4:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 27 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: other nucleic acid
(A) DESCRIPTION: /desc = "Oligonucleotide"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4: CGTTCTAAGG ATTACGTCGA ACCATTG 27
(2) INFORMATION FOR SEQ ID NO: 5:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 9 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: other nucleic acid
(A) DESCRIPTION: /desc = "Oligonucleotide"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5: GCAAGATTC (2) INFORMATION FOR SEQ ID NO: 6:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 9 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: other nucleic acid
(A) DESCRIPTION: /desc = "Oligonucleotide"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6: CTAATGCAG (2) INFORMATION FOR SEQ ID NO: 7:
(i) SEQUENCE CHARACTERISTICS :
(A) LENGTH: 9 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: other nucleic acid
(A) DESCRIPTION: /desc = "Oligonucleotide"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7: CTTGGAAAC

Claims

1. A method for determining the presence and location of a mismatch in a target sequence, which comprises contacting the target sequence with first, second and third oligonucleotides capable of hybridising to the natural sequence, in juxtaposition, wherein the oligonucleotides are respectively labelled with first, second and third markers capable of absorption at first, second and third wavelengths and emission at first, second and third wavelengths, such that there is resonance between either or each of first and second markers and between second and third markers, and observing the presence or absence of resonance.
2. A method according to claim 1, wherein the second oligonucleotide comprises two second markers resonating with the first and third markers respectively.
3. A method according to claim 1 or claim 2, wherein one or more markers is a fluorescent moiety.
4. A method according to any preceding claim, wherein the one or more markers are selected from 5' -carboxyrhoda ine-
X, 5 ' -carboxytetramethylrhodamine, N,N,N' ,N'-tetramethyl-6- carboxyrhodamine, 2 ' , 7 ' -dimethoxy-4 ' , 5 ' -dichloro-6- carboxyfluorescein and 5 '-carboxyfluorescein.
5. A method according to any preceding claim, wherein the oligonucleotides each comprise 8-13 bases.
6. A method according to any preceding claim, wherein adjacent markers are separated by at least 7 bases.
7. A method according to any preceding claim, wherein the target or the first, second and third oligonucleotides are immobilised.
8. A kit comprising, in separate compartments, first, second and third markers defined in any of claims 1 to 5.
PCT/GB1998/001138 1997-04-21 1998-04-20 Dna mutation mapping by multiple energy transfer interactions WO1998048048A2 (en)

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Application Number Priority Date Filing Date Title
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GB9707996.6 1997-04-21
GBGB9707996.6A GB9707996D0 (en) 1997-04-21 1997-04-21 Screening

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Cited By (9)

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Publication number Priority date Publication date Assignee Title
WO1999041607A2 (en) * 1998-02-14 1999-08-19 Gmd Forschungszentrum Informationstechnik Gmbh Fluorescent energy for elucidating the 3-d structure of biological macromolecules
WO2000018965A1 (en) * 1998-09-30 2000-04-06 University Of Utah Research Foundation Multiplex genotyping using fluorescent hybridization probes
DE19850593A1 (en) * 1998-11-03 2000-05-04 Biochip Technologies Gmbh DNA hybridization assay comprises detecting fluorescent resonance energy transfer between fluorochromes on different DNA strands
WO2000051056A2 (en) * 1999-02-22 2000-08-31 Vialogy Corporation Method and apparatus for interpreting dna microarray patterns
WO2000052625A2 (en) * 1999-02-22 2000-09-08 Vialogy Corporation Method and apparatus for analyzing hybridized biochip patterns using resonance interactions
WO2001092564A1 (en) * 2000-05-29 2001-12-06 The Walter And Eliza Hall Institute Of Medical Research A method for determining the likelihood that a test polynucleotide sequence differs from a driver polynucleotide
WO2002095057A2 (en) * 2001-05-24 2002-11-28 Genospectra, Inc. Pairs of nucleic acid probes with interactive signaling moieties and nucleic acid probes with enhanced hybridization efficiency and specificity
WO2005050182A1 (en) * 2003-11-19 2005-06-02 Dimerix Biosciences Pty Ltd Resonance energy transfer assay system for multi-component detection
US8484000B2 (en) 2004-09-02 2013-07-09 Vialogy Llc Detecting events of interest using quantum resonance interferometry

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WO1993009128A1 (en) * 1991-11-07 1993-05-13 Nanotronics, Inc. Hybridization of polynucleotides conjugated with chromophores and fluorophores to generate donor-to-donor energy transfer system
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WO1996025518A1 (en) * 1995-02-17 1996-08-22 The Society For Techno-Innovation Of Agriculture, Foresty And Fisheries Probe for use in nucleic acid analysis and detecting method

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EP0229943B1 (en) * 1985-12-23 1991-09-04 Molecular Biosystems, Inc. Fluorescent stokes shift probes for polynucleotide hybridization assays
WO1993009128A1 (en) * 1991-11-07 1993-05-13 Nanotronics, Inc. Hybridization of polynucleotides conjugated with chromophores and fluorophores to generate donor-to-donor energy transfer system
EP0601889A2 (en) * 1992-12-10 1994-06-15 Maine Medical Center Research Institute Nucleic acid probes
WO1996025518A1 (en) * 1995-02-17 1996-08-22 The Society For Techno-Innovation Of Agriculture, Foresty And Fisheries Probe for use in nucleic acid analysis and detecting method

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Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6713256B1 (en) * 1998-02-14 2004-03-30 Fraunhofer Gesellschaft zur Förderung der angewandten Forschung e.V. Fluorescent energy transfer mediated chemical activation (fetma) for the elucidation of the three-dimensional structure of biomacromolecules
WO1999041607A3 (en) * 1998-02-14 1999-12-09 Gmd Gmbh Fluorescent energy for elucidating the 3-d structure of biological macromolecules
WO1999041607A2 (en) * 1998-02-14 1999-08-19 Gmd Forschungszentrum Informationstechnik Gmbh Fluorescent energy for elucidating the 3-d structure of biological macromolecules
WO2000018965A1 (en) * 1998-09-30 2000-04-06 University Of Utah Research Foundation Multiplex genotyping using fluorescent hybridization probes
DE19850593A1 (en) * 1998-11-03 2000-05-04 Biochip Technologies Gmbh DNA hybridization assay comprises detecting fluorescent resonance energy transfer between fluorochromes on different DNA strands
WO2000051056A2 (en) * 1999-02-22 2000-08-31 Vialogy Corporation Method and apparatus for interpreting dna microarray patterns
WO2000052625A2 (en) * 1999-02-22 2000-09-08 Vialogy Corporation Method and apparatus for analyzing hybridized biochip patterns using resonance interactions
WO2000051056A3 (en) * 1999-02-22 2000-12-07 Vialogy Corp Method and apparatus for interpreting dna microarray patterns
WO2000052625A3 (en) * 1999-02-22 2001-01-25 Vialogy Corp Method and apparatus for analyzing hybridized biochip patterns using resonance interactions
WO2001092564A1 (en) * 2000-05-29 2001-12-06 The Walter And Eliza Hall Institute Of Medical Research A method for determining the likelihood that a test polynucleotide sequence differs from a driver polynucleotide
US8666669B2 (en) 2000-05-29 2014-03-04 Genera Biosystems Limited Method for determining the likelihood that a test polynucleotide sequence differs from a driver polynucleotide
WO2002095057A3 (en) * 2001-05-24 2003-02-20 Genospectra Inc Pairs of nucleic acid probes with interactive signaling moieties and nucleic acid probes with enhanced hybridization efficiency and specificity
WO2002095057A2 (en) * 2001-05-24 2002-11-28 Genospectra, Inc. Pairs of nucleic acid probes with interactive signaling moieties and nucleic acid probes with enhanced hybridization efficiency and specificity
WO2005050182A1 (en) * 2003-11-19 2005-06-02 Dimerix Biosciences Pty Ltd Resonance energy transfer assay system for multi-component detection
JP2007511226A (en) * 2003-11-19 2007-05-10 ディメリックス バイオサイエンス ピーティーワイ リミテッド Resonant energy transfer assay system for multiple component detection
US8484000B2 (en) 2004-09-02 2013-07-09 Vialogy Llc Detecting events of interest using quantum resonance interferometry

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AU7064198A (en) 1998-11-13
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