Method of sequencing of genomes by hybridization of oligonucleot

Details Method of sequencing of genomes by hybridization of oligonucleot Method of sequencing of genomes by hybridization of oligonucleot

1991-06-18

1993-04-13

Moskowitz, Margaret

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

436501, 436 94, 935 77, 935 78, 536 2432, 536 231, C12Q 168, C07H 1512, G01N 33566, G01N 3348

Patent

active

052022315

ABSTRACT:
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.

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., Nuc. Acids Res. 13(22):8083-8091 (1985).
Maniatis et al., Molecular Cloning A Laboratory Manual Cold Spring Harbor Lab., N.Y. 1982.
Poustka et al., Cold Spring Harb. Symp. Quant. Biol. LI:131-139 (1986).

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