Microchip method for the enrichment of specific DNA sequences
First Claim
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1. 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.
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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.
120 Citations
19 Claims
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1. A method for enriching specific genetic sequences, comprising:
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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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. A method for enriching specific genetic sequences contained in genetic material, comprising:
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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. - View Dependent Claims (13, 14, 15, 16, 17, 18, 19)
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Specification