US5492806A
Expired / Lapsed
US
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Method of determining an ordered sequence of subfragments of a nucleic acid fragment by hybridization of oligonucleotide probes
Assignee: Hyseq Inc
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Filing: 1993-04-12
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Granted: 1996-02-20
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Expires: 2013-04-12
Abstract
Abstract The sequence of a given nucleic acid fragment is read by the hybridization and assembly of positively hybridizing exactly complementary oligonucleotide probes through overlapping subfragments. By simultaneous hybridization of nucleic acid subfragments bound onto a filter, representing single-stranded phage vector with a cloned insert, with about 50,000 to 100,000 groups of probes, the main type of which is (A,T,C,G)(A,T,C,G)N8(A,T,C,G), information for computer determination of a sequence of DNA having the complexity of a mammalian genome are obtained in one step. To obtain a maximally completed sequence, three libraries cloned into the phage vector, M13, are used. The process can be easily and entirely robotized for factory reading of complex genomic fragments or DNA molecules.
Inventors
- 👤 Radoje T. Drmanac
- 👤 Radomir B. Crkvenjakov
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-28 21:25
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6869; C12Q1/6874; C12Q1/6813; C12Q1/6827; Y; Y10; Y10T; Y10T436/00; Y10T436/14; Y10T436/142222; Y10T436/143333
Claims Analysis
Total Claims: 2
Claim 1
Independent Claim
179 words
A method for determining an ordered sequence of one or more subfragments of a nucleic acid fragment by ordering oligonucleotide probe sequences which are complementary to subfragments of said nucleic acid fragment, comprising the steps of: (a) contacting said nucleic acid fragment with a set of oligonucleotide probes, each having a unique internal portion selected from the group consisting of 5-mers, 6-mers, 7-mers, and 8-mers, which may be in tandem, under conditions which distinguish a subset of oligonucleotide probes which are exactly complementary to one or more subfragments of said nucleic acid fragment from oligonucleotide probes which are not exactly complementary to one or more subfragments of said nucleic acid fragment; (b) detecting the subset of oligonucleotide probes which are exactly complementary to one or more subfragments of said nucleic acid fragment; and (c) ordering said subset of oligonucleotide probes by compiling overlapping sequences of said subset of oligonucleotide probes which are exactly complementary to one or more subfragments of said nucleic acid fragment, thereby determining the ordered sequence of one or more subfragments of a nucleic acid fragment.
Claim 2
Independent Claim
179 words
A method for determining an ordered sequence of one or more subfragments of a nucleic acid fragment by ordering oligonucleotide probe sequences which are complementary to subfragments of said nucleic acid fragment, comprising the steps of: (a) contacting said nucleic acid fragment with a set of oligonucleotide probes, each having a unique internal portion selected from the group consisting of 5-mers, 6-mers, 7-mers, and 8-mers, which may be in tandem, under conditions which distinguish a subset of oligonucleotide probes which are exactly complementary to one or more subfragments of said nucleic acid fragment from oligonucleotide probes which are not exactly complementary to one or more subfragments of said nucleic acid fragment; (b) detecting the subset of oligonucleotide probes which are exactly complementary to one or more subfragments of said nucleic acid fragment; and (c) ordering said subset of oligonucleotide probes by compiling overlapping sequences of said subset of oligonucleotide probes which are exactly complementary to one or more subfragments of said nucleic acid fragment, thereby determining the ordered sequence of one or more subfragments of a nucleic acid fragment.
References Cited (Backward Citations)
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1999-02-19 | 1999-08-21 | 2000-02-20 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2003-02-19 | 2003-08-21 | 2004-02-20 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2007-02-19 | 2007-08-21 | 2008-02-20 | $7700.00 | Window Closed / Lapsed |