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US5851772A
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Microchip method for the enrichment of specific DNA sequences
Assignee: UChicago Argonne LLC
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Filing: 1996-01-29
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Granted: 1998-12-22
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Expires: 2016-01-29
Abstract
Abstract A method for enriching specific genetic material sequences is provided, whereby oligonucleotide molecules complementary to the desired genetic material is first used to isolate the genetic material from a first source of genomic material. Then the genetic material is used as a label to isolate similar genetic sequences from other sources.
Inventors
- 👤 Andrei Darievich Mirzabekov
- 👤 Yuri Petrovich Lysov
- 👤 Valentine Vladimirovich Shick
- 👤 Svetlana Alekseevna Dubiley
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 18:31
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6811; C12Q1/6813; C12Q1/6834; C12Q1/6837; B; B01; B01J; B01J2219/00; B01J2219/00274; B01J2219/00277; B01J2219/00497; B01J2219/00527; B01J2219/00529; B01J2219/00583; B01J2219/00603; B01J2219/00605; B01J2219/00608; B01J2219/0061; B01J2219/00612; B01J2219/00614; B01J2219/00617; B01J2219/00619; B01J2219/00623; B01J2219/00626; B01J2219/00639; B01J2219/00644; B01J2219/00659; B01J2219/00662; B01J2219/00718; B01J2219/0072; B01J2219/00722; C40; C40B; C40B40/00; C40B40/04; C40B40/06
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
174 words
A method for enriching specific genetic sequences, comprising: a) immobilizing a first set of oligonucleotide molecules at a plurality of positions on a first gel substrate, said first set of oligonucleotide molecules synthesized to complement base sequences of the genetic sequences; b) contacting genetic material with said first set of oligonucleotides, said genetic material containing the specific genetic sequences, so as to allow formation of first duplexes of said first set of oligonucleotides and fragments of the specific genetic sequences; c) simultaneously isolating the fragments of the specific genetic material sequences from the duplexes formed via diffusion; d) labeling the now-isolated fragments with a fluorochrome; e) simultaneously contacting each now-labeled fragment with a second separate set of sequencing oligonucleotide molecules contained on a sequencing gel substrate, said sequencing set of oligonucleotide molecules which are complementary to subsequence regions of each now-labelled fragment, so as to allow for second duplexes to form between the subsequence regions and said sequencing set of oligonucleotide molecules; and f) isolating the subsequence regions, thereby enriching the specific genetic sequences.
Claim 2
Independent Claim
174 words
A method for enriching specific genetic sequences, comprising: a) immobilizing a first set of oligonucleotide molecules at a plurality of positions on a first gel substrate, said first set of oligonucleotide molecules synthesized to complement base sequences of the genetic sequences; b) contacting genetic material with said first set of oligonucleotides, said genetic material containing the specific genetic sequences, so as to allow formation of first duplexes of said first set of oligonucleotides and fragments of the specific genetic sequences; c) simultaneously isolating the fragments of the specific genetic material sequences from the duplexes formed via diffusion; d) labeling the now-isolated fragments with a fluorochrome; e) simultaneously contacting each now-labeled fragment with a second separate set of sequencing oligonucleotide molecules contained on a sequencing gel substrate, said sequencing set of oligonucleotide molecules which are complementary to subsequence regions of each now-labelled fragment, so as to allow for second duplexes to form between the subsequence regions and said sequencing set of oligonucleotide molecules; and f) isolating the subsequence regions, thereby enriching the specific genetic sequences.
Claim 3
Dependent Claim
23 words
The method as recited in claim 1 wherein the genetic material is selected from the group consisting of deoxyribonucleic acid and ribonucleic acid.
Claim 4
Dependent Claim
31 words
The method as recited in claim 1 wherein the first set and second set of oligonucleotide molecules contain bases selected from the group consisting of guanine, cytosine, adenine, thymine and uracil.
Claim 5
Dependent Claim
24 words
The method as recited in claim 1 wherein the first set of oligonucleotide molecules consist of different base sequences and are of equal length.
Claim 6
Dependent Claim
20 words
The method as recited in claim 1 wherein the oligonucleotide molecules range in length from approximately 6 and 12 bases.
Claim 7
Dependent Claim
28 words
The method as recited in claim 1 wherein the fluorochrome is selected from the group consisting of tetramethylrodamine, fluoresceine, Texas Red, Cascade Blue, Rhodamine, HEX, and combinations thereof.
Claim 8
Dependent Claim
26 words
The method as recited in claim 1 wherein the substrate is selected from the group consisting of polyacrylamide, polyamide, agarose, silicate glass matrix, and combinations thereof.
Claim 9
Dependent Claim
19 words
The method as recited in claim 1 wherein the second set of oligonucleotide molecules further comprises a universal base.
Claim 10
Dependent Claim
24 words
The method as recited in claim 8 wherein the universal base is selected from the group consisting of 5-nitroindole, 3-nitropyrrole, inosine, and combinations thereof.
Claim 11
Dependent Claim
34 words
The method as recited in claim 1 wherein said genetic material is fragmented into lengths of between approximately 250 and 30,000 bases prior to contacting said genetic material with said first set of oligonucleotides.
Claim 12
Dependent Claim
35 words
The method as recited in claim 1 wherein said now isolated fragments are fragmented into lengths of between approximately 10 to 50 bases prior to contacting said now-isolated fragments with the second set of oligonucleotides.
Claim 13
Independent Claim
209 words
A method for enriching specific genetic sequences contained in genetic material, comprising: a) fractionating genetic material to lengths of between approximately 250 and 30,000 bases; b) labeling the now fractionated genetic material with a fluorochrome; c) immobilizing a first set of oligonucleotide molecules at a plurality of positions on a first porous substrate, said first set of oligonucleotide molecules synthesized to complement base sequences of the genetic material; d) contacting the now-labeled genetic material with said first set of oligonucleotides so as to allow formation of duplexes of said first set of oligonucleotides and the specific genetic sequences; e) simultaneously isolating each of the specific genetic material sequences from the duplexes formed via diffusion; f) fragmenting the now-isolated specific genetic material sequences into lengths of between approximately 10 and 50 bases; g) labeling the now-isolated specific genetic material with a fluorochrome; h) simultaneously contacting each now-labeled fragment with a separate set of sequencing oligonucleotide molecules contained on a sequencing gel substrate, said sequencing set of oligonucleotide molecules which are complementary to subsequence regions of each now-labelled fragment, so as to allow for second duplexes to form between the subsequence regions and said sequencing set of oligonucleotide molecules; and i) isolating the subsequence regions thereby enriching the specific genetic sequences.
Claim 14
Independent Claim
209 words
A method for enriching specific genetic sequences contained in genetic material, comprising: a) fractionating genetic material to lengths of between approximately 250 and 30,000 bases; b) labeling the now fractionated genetic material with a fluorochrome; c) immobilizing a first set of oligonucleotide molecules at a plurality of positions on a first porous substrate, said first set of oligonucleotide molecules synthesized to complement base sequences of the genetic material; d) contacting the now-labeled genetic material with said first set of oligonucleotides so as to allow formation of duplexes of said first set of oligonucleotides and the specific genetic sequences; e) simultaneously isolating each of the specific genetic material sequences from the duplexes formed via diffusion; f) fragmenting the now-isolated specific genetic material sequences into lengths of between approximately 10 and 50 bases; g) labeling the now-isolated specific genetic material with a fluorochrome; h) simultaneously contacting each now-labeled fragment with a separate set of sequencing oligonucleotide molecules contained on a sequencing gel substrate, said sequencing set of oligonucleotide molecules which are complementary to subsequence regions of each now-labelled fragment, so as to allow for second duplexes to form between the subsequence regions and said sequencing set of oligonucleotide molecules; and i) isolating the subsequence regions thereby enriching the specific genetic sequences.
Claim 15
Dependent Claim
23 words
The method as recited in claim 12 wherein the genetic material is selected from the group consisting of deoxyribonucleic acid and ribonucleic acid.
Claim 16
Dependent Claim
30 words
The method as recited in claim 12 wherein the first and second set of oligonucleotide molecules contain bases selected from the group consisting of guanine, cytosine, adenine, thymine and uracil.
Claim 17
Dependent Claim
24 words
The method as recited in claim 12 wherein the first set of oligonucleotide molecules consist of different base sequences and are of equal length.
Claim 18
Dependent Claim
19 words
The method as recited in claim 12 wherein the length of the oligonucleotides is between 6 and 12 bases.
Claim 19
Dependent Claim
33 words
The method as recited in claim 12 wherein the genetic material is labeled with a fluorochrome selected from the group consisting of tetramethylrodamine, fluoresceine, Texas Red, Cascade Blue, Rhodamine, HEX, and combinations thereof.
Claim 20
Dependent Claim
28 words
The method as recited in claim 12 wherein the substrate is selected from the group consisting of polyacrylamide gel, polyamide gel, agarose, silicate glass matrix and combinations thereof.
Claim 21
Dependent Claim
20 words
The method as recited in claim 12 wherein the second set of oligonucleotides are further comprised of a universal base.
References Cited (Backward Citations)
US5434049A
Cited
US5552278A
Cited
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