Apparatus for determining base sequence of nucleic acid
US6242193A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | May 9, 2000 |
| Grant date | Jun 5, 2001 |
| Priority date | — |
| Expiry date | May 9, 2020 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N21/6486
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
A single molecule of single-stranded sample DNA (7) having a bead (5) at one end and a magnetic bead (6) at the other end is extended and fixed in the field of view of a fluorescent microscope by using a magnetic force (11) and a laser trap (3), and a primer (8) is bonded thereto, followed by elongation reaction (10) using polymerase. Only a single chemically modified nucleotide (9) labeled with at least one fluorophore which varies depending on the kind of the base is incorporated. Only the single fluorophore incorporated is measured as a fluorescence-microscopic image by evanescent irradiation (13) with exciting laser beams, and the kind of the base is determined from the kind of the fluorophore. The fluorophore labeling the nucleotide incorporated is released by evanescent irradiation (13) with ultraviolet laser beams (2), and the next nucleotide is incorporated. DNA sequencing is carried out by repeating the above procedure. The base sequence determination can be carried out by using the single DNA molecule, so that a DNA base sequence of hundreds kilos or more bases can be efficiently determined.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.