Method of determining an ordered sequence of subfragments of a n

Details Method of determining an ordered sequence of subfragments of a n Method of determining an ordered sequence of subfragments of a n

1993-04-12

1996-02-20

Zitomer, Stephanie W.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving virus or bacteriophage

435 6, 935 77, 935 78, C12Q 170, C12Q 168

Patent

active

054928060

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.

REFERENCES:
patent: 4562159 (1985-12-01), Shafritz
patent: 4613566 (1986-09-01), Potter
patent: 4683202 (1987-07-01), Mullis
patent: 4849334 (1989-07-01), Lorincz
Chan et al., "Detection of subpicogram quantities of specific DNA sequences on blot hybridization with biotinylated probes," Nucleic Acids Research, 13(22):8083-8091 (1985).
Maniatis et al., "Analysis of Recombinant DNA Clones," Molecular Cloning A Labratory Manual, p. 388 (Cold Spring Harbor Labratory) (1982).
Poustka et al., "Molecular Approaches to Mammalian Genetics," Cold Spring Harb. Symp. Quant. Biol., LI:131-139 (1986).

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