Method of determining an ordered sequence of subfragments of a nucleic acid fragment by hybridization of oligonucleotide probes
US5492806A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Apr 12, 1993 |
| Grant date | Feb 20, 1996 |
| Priority date | — |
| Expiry date | Apr 12, 2013 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY10T436/143333
- WIPO fieldBiotechnology
- WIPO sectorChemistry
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.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.