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US5849486A Expired / Lapsed US ⏱️ 0.0 years remaining

Methods for hybridization analysis utilizing electrically controlled hybridization

Assignee: Gamida for Life BV
•
Filing: 1995-09-27
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Granted: 1998-12-15
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Expires: 2015-09-27
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Abstract

Abstract A system for performing molecular biological diagnosis, analysis and multistep and multiplex reactions utilizes a selfaddressable, selfassembling microelectronic system for actively carrying out controlled reactions in microscopic formats. Preferably, a fluidic system flow a sample across an active area of the biochip, increasing diagnostic efficiency. Preferably, the fluidic system includes aflow cell having a window. Pulsed activation of the electrodes of the biochip are advantageously employed with the fluidic system, permitting more complete sampling of the materials within the biological sample. An improved detection system utilizes a preferably coaxially oriented excitation fiber, such as a fiber optic, disposed within a light guide, such as a liquid light guide. In this way, small geometry systems may be fluorescently imaged. A highly automated DNA diagnostic system results. Perturbation of the fluorescence signal during electronic denaturation is detailed and analyzed for analytical and diagnostic purposes. Such fluorescence perturbation information is combined with other information to provide improved analysis. DNA fingerprinting uses hybridizing DNA fragments of a given length and a capture sequence at a test site and then determining the level of reverse bias necessary to affect the hybridization, such as to dehybridize, and determine the length of the DNA.

Inventors

  • 👤 Michael James Heller
  • 👤 James Patrick O'Connell
  • 👤 Robert David Juncosa
  • 👤 Ronald George Sosnowski
  • 👤 Thomas Ratcliffe Jackson

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-11-07 05:41
CPC Classifications
G; G11; G11C; G11C13/00; G11C13/0002; G11C13/0009; G11C13/0014; G11C13/0019; B; B01; B01J; B01J19/00; B01J19/0046; B01J19/0093; B01L; B01L3/00; B01L3/50; B01L3/502; B01L3/5027; B01L3/502761; B01L3/508; B01L3/5085; B82; B82Y; B82Y10/00; B82Y5/00; C; C07; C07H; C07H21/00; C07K; C07K1/00; C07K1/04; C07K1/045; C07K1/047; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/6825; C12Q1/6834; C12Q1/6837; G11C19/00; H; H10; H10W; H10W90/00; B01J2219/00; B01J2219/00274; B01J2219/00277; B01J2219/00279; B01J2219/00306; B01J2219/00313; B01J2219/00315; B01J2219/00317; B01J2219/00497; B01J2219/00527; B01J2219/00583; B01J2219/00585; B01J2219/0059; B01J2219/00596; B01J2219/00603; B01J2219/00605; B01J2219/00612; B01J2219/00614; B01J2219/00621; B01J2219/00623; B01J2219/00626; B01J2219/00632; B01J2219/00637; B01J2219/00639; B01J2219/00644; B01J2219/00653; B01J2219/00659; B01J2219/0068; B01J2219/00686; B01J2219/00689; B01J2219/00709; B01J2219/00713; B01J2219/00718; B01J2219/0072; B01J2219/00722; B01J2219/00725; B01J2219/00731; B01J2219/00781; B01J2219/00819; B01J2219/00824; B01J2219/00828; B01J2219/00851; B01J2219/00853; B01J2219/0095; B01J2219/00952; B01J2219/00968; B01J2219/0097; B01L2200/00; B01L2200/02; B01L2200/025; B01L2200/14; B01L2200/143; B01L2300/00; B01L2300/06; B01L2300/0627; B01L2300/0636; B01L2300/0645; B01L2300/08; B01L2300/0809; B01L2300/0816; B01L2300/0832; B01L2300/0838; B01L2300/0861; B01L2300/0877; B01L2400/00; B01L2400/04; B01L2400/0403; B01L2400/0415; B01L2400/0421; B01L9/00; B01L9/52; B01L9/527; C07B; C07B2200/00; C07B2200/11; C40; C40B; C40B40/00; C40B40/04; C40B40/06; C40B40/10; C40B40/12; C40B60/00; C40B60/14; G11C13/04; H10W72/00; H10W72/50; H10W72/531; H10W72/5363; H10W72/551; H10W72/552; H10W72/5522; H10W72/851; H10W72/874; H10W72/884; H10W90/701; H10W90/731; H10W90/734; H10W90/751; H10W90/754

Claims Analysis

Total Claims: 39
Claim 1
Independent Claim 83 words
A method for hybridization analysis between a sample and a probe, the analysis utilizing an electronic stringency control device, comprising the steps of: providing the sample and probe with a fluorescent label under hybridization conditions on the electronic stringency control device, forming a fluorescently labelled hybridization product, monitoring the fluorescence from the hybridization product, subjecting the hybridization product to varying electrophoretic force, and analyzing the fluorescent signal from the flourescently labeled hybridization product for variance as a function of said varying electrophoretic force.
Claim 2
Independent Claim 83 words
A method for hybridization analysis between a sample and a probe, the analysis utilizing an electronic stringency control device, comprising the steps of: providing the sample and probe with a fluorescent label under hybridization conditions on the electronic stringency control device, forming a fluorescently labelled hybridization product, monitoring the fluorescence from the hybridization product, subjecting the hybridization product to varying electrophoretic force, and analyzing the fluorescent signal from the flourescently labeled hybridization product for variance as a function of said varying electrophoretic force.
Claim 3
Dependent Claim 18 words
The method for hybridization analysis of claim 1 wherein the fluorescence is analyzed for the fluorescent perturbation value.
Claim 4
Dependent Claim 19 words
The method for hybridization analysis of claim 2 wherein the fluorescence perturbation value is measured for the onset value.
Claim 5
Dependent Claim 18 words
The method for hybridization analysis of claim 2 wherein the fluorescence perturbation value is measured for its height.
Claim 6
Dependent Claim 18 words
The method for hybridization analysis of claim 2 wherein the fluorescence perturbation value is measured for the slope.
Claim 7
Dependent Claim 20 words
The method for hybridization analysis of claim 1 wherein the fluorescence is analyzed for the power level of the perturbation.
Claim 8
Dependent Claim 57 words
The method for hybridization analysis of claim 1 further including the steps of: determining a second measure of hybridization between the sample and the probe, and combining the information obtained by the first analysis including the step of subjecting the hybridization product to the varying electrophoretic force in the second measure to provide a indication of hybridization.
Claim 9
Dependent Claim 21 words
The method for hybridization analysis of claim 7 wherein the second measure of hybridization includes determination of the electronic melting point.
Claim 10
Dependent Claim 21 words
The method for hybridization analysis of claim 1 wherein the fluorescent label is placed in proximity to the initial denaturation site.
Claim 11
Independent Claim 22 words
The method for hybridization analysis of claims 1 or 9 wherein the fluorescent label is intercalated adjacent a single base mismatch site.
Claim 12
Dependent Claim 15 words
The method for hybridization analysis of claim 10 wherein the fluorescent label is ethidium bromide.
Claim 13
Dependent Claim 14 words
The method for hybridization analysis of claim 10 wherein the fluorescent label is acridine.
Claim 14
Independent Claim 31 words
The method for hybridization analysis of claims 1 or 7 wherein the electrophoretic force is in an amount less than is necessary to effect dehybridization of the sample and the probe.
Claim 15
Dependent Claim 19 words
The method for hybridization analysis of claim 1 wherein the hybridization product is subject to an oscillating electrophoretic force.
Claim 16
Dependent Claim 21 words
The method for hybridization analysis of claim 7 wherein the fluorescent label is placed in proximity to the initial denaturation site.
Claim 17
Dependent Claim 30 words
The method for hybridization analysis of claim 13 wherein the electrophoretic force is in an amount less than is necessary to effect complete denaturation of the sample and the probe.
Claim 18
Dependent Claim 34 words
The method for hybridization analysis of claim 14 where the fluorescent signal from a mismatched sample and probe exhibits a relatively greater response to the oscillating electrophoretic force relative to a matched hybridization product.
Claim 19
Dependent Claim 16 words
The method for hybridization analysis of claim 1 wherein the probe is longer than a 20-mer.
Claim 20
Dependent Claim 18 words
The method for hybridization analysis of claim 1 further including the step of averaging of the fluorescent signal.
Claim 21
Independent Claim 69 words
A method for hybridization analysis between a sample and a probe comprising the steps of: providing the sample and probe with a fluorescent label under hybridization conditions, forming a fluorescently labeled hybridization product, monitoring the fluorescence from the hybridization product, subjecting the hybridization product to varying electrophoretic force, and analyzing the fluorescent signal for variance as a function of said varying electrophoretic force to determine the fluorescence perturbation value.
Claim 22
Independent Claim 69 words
A method for hybridization analysis between a sample and a probe comprising the steps of: providing the sample and probe with a fluorescent label under hybridization conditions, forming a fluorescently labeled hybridization product, monitoring the fluorescence from the hybridization product, subjecting the hybridization product to varying electrophoretic force, and analyzing the fluorescent signal for variance as a function of said varying electrophoretic force to determine the fluorescence perturbation value.
Claim 23
Dependent Claim 19 words
The method for hybridization analysis of claim 20 wherein the fluorescence perturbation value is measured for the onset value.
Claim 24
Dependent Claim 18 words
The method for hybridization analysis of claim 20 wherein the fluorescence perturbation value is measured for its height.
Claim 25
Dependent Claim 18 words
The method for hybridization analysis of claim 20 wherein the fluorescence perturbation value is measured for the slope.
Claim 26
Dependent Claim 20 words
The method for hybridization analysis of claim 20 wherein the fluorescence is analyzed for the power level of the perturbation.
Claim 27
Dependent Claim 57 words
The method for hybridization analysis of claim 20 further including the steps of: determining a second measure of hybridization between the sample and the probe, and combining the information obtained by the first analysis including the step of subjecting the hybridization product to the varying electrophoretic force in the second measure to provide a indication of hybridization.
Claim 28
Dependent Claim 21 words
The method for hybridization analysis of claim 25 wherein the second measure of hybridization includes determination of the electronic melting point.
Claim 29
Dependent Claim 21 words
The method for hybridization analysis of claim 20 wherein the fluorescent label is placed in proximity to the initial denaturation site.
Claim 30
Dependent Claim 20 words
The method for hybridization analysis of claim 20 wherein the fluorescent label is intercalated adjacent a single based mismatch site.
Claim 31
Independent Claim 17 words
The method for hybridization analysis of claims 27 or 28 wherein the fluorescent label is ethidium bromide.
Claim 32
Independent Claim 16 words
The method for hybridization analysis of claims 27 or 28 wherein the fluorescent label is acridine.
Claim 33
Dependent Claim 29 words
The method for hybridization analysis of claim 20 wherein the electrophoretic force is in an amount less than is necessary to effect dehybridization of the sample and the probe.
Claim 34
Dependent Claim 19 words
The method for hybridization analysis of claim 20 wherein the hybridization product is subject to an oscillating electrophoretic force.
Claim 35
Dependent Claim 21 words
The method for hybridization analysis of claim 25 wherein the fluorescent label is placed in proximity to the initial denaturation site.
Claim 36
Dependent Claim 30 words
The method for hybridization analysis of claim 31 wherein the electrophoretic force is in an amount less than is necessary to effect complete denaturation of the sample and the probe.
Claim 37
Dependent Claim 34 words
The method for hybridization analysis of claim 32 where the fluorescent signal from a mismatch sample and probe exhibits a relatively greater response to the oscillating electrophoretic force relative to a matched hybridization product.
Claim 38
Dependent Claim 16 words
The method for hybridization analysis of claim 20 wherein the probe is longer than a 20-mer.
Claim 39
Dependent Claim 18 words
The method for hybridization analysis of claim 20 further including the step of averaging of the fluorescent signal.

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