Microsystem for rapid DNA sequencing
US5846727A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | May 29, 1997 |
| Grant date | Dec 8, 1998 |
| Priority date | — |
| Expiry date | May 29, 2017 |
Classification
- Technology area (CPC B)Performing Operations; Transporting
- CPC primaryB01L2400/0421
- WIPO fieldChemical engineering
- WIPO sectorChemistry
Abstract
A system is disclosed for the rapid and cost-effective sequencing of DNA. There are three principal components of the system: (1) a microreactor, which prepares DNA sequencing "ladders" using solid-phase techniques, preferably in capillary tubes whose volumes are on the order of 10-1000 nanoliters, preferably 10-200 nanoliters; (2) a microfabricated electrophoresis capillary separation unit; and (3) a fluorescence detector with single-mode optical fibers interfaced directly to the electrophoresis capillary. The system is suitable for a highly multiplexed, automated DNA sequencing device Typical. steps in sequencing are as follows: (1) PCR amplification of a DNA template in microtiter dishes using labelled primers, e.g., primers labelled with biotin; (2) immobilizing the labelled PCR products on the walls of one or more capillary tubes having volumes on the order of 10-200 nanoliters; (3) preparing nanoliter quantities of labelled Sanger extension products of the amplified DNA; (4) purifying the oligonucleotide sequencing ladders; (5) high speed electrophoretic separation of the sequencing ladders; and (6) near-infrared, laser-induced fluorescence detection of the oligonucleotides. …
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.