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| 국가/구분 | United States(US) Patent 등록 |
|---|---|
| 국제특허분류(IPC7판) |
|
| 출원번호 | US-0723712 (1991-06-18) |
| 우선권정보 | YU-0000570 (1987-04-01) |
| 발명자 / 주소 |
|
| 인용정보 | 피인용 횟수 : 534 인용 특허 : 0 |
The conditions under which oligonucleotides hybridize only with entirely homologous sequences are recognized. The sequence of a given DNA fragment is read by the hybridization and assembly of positively hybridizing probes through overlapping portions. By simultaneous hybridization of DNA molecules a
The conditions under which oligonucleotides hybridize only with entirely homologous sequences are recognized. The sequence of a given DNA fragment is read by the hybridization and assembly of positively hybridizing probes through overlapping portions. By simultaneous hybridization of DNA molecules applied as dots and bound onto a filter, representing single-stranded phage vector with the 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, bacteriophage are used: with the 0.5 kb and 7 kbp insert consisting of two sequences, with the average distance in genomic DNA of 100 kbp. For a million bp of genomic DNA, 25,000 subclones of the 0.5 kbp are required as well as 700 subclones 7 kb long and 170 jumping subclones. Subclones of 0.5 kb are applied on a filter in groups of 20 each, so that the total number of samples is 2,120 per million bp. The process can be easily and entirely robotized for factory reading of complex genomic fragments or DNA molecules.
A method of sequencing a nucleic acid fragment, comprising: (a) contacting a nucleic acid fragment with a set of oligonucleotide probes of predetermined sequence and length under hybridization conditions that allow differentiation between (i) those probes of said set which are exactly complementary
A method of sequencing a nucleic acid fragment, comprising: (a) contacting a nucleic acid fragment with a set of oligonucleotide probes of predetermined sequence and length under hybridization conditions that allow differentiation between (i) those probes of said set which are exactly complementary to part of said fragment and (ii) those probes of said set which are not exactly complementary to part of said fragment; (b) detecting those probes of said set which are exactly complementary to part of said fragment; and (c) determining the sequence of said fragment from the subset of those probes of said set which are detected as exactly complementary to part of said fragment by compiling their overlapping sequences.
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