Shimadzu Corporation

Hidehiko Fujii

Kunihiko Ohkubo

Disclosed herein is an on-line base sequencing apparatus, which is provided with electrophoresis tubes having a sheath flow part in each lower end thereof, which a sulfuric acid solution of pH 1.0 is continuously fed to form a sheath flow. The portions close to the outlets provided in the lower ends are irradiated with light which is close to 260 nm. In order to detect fluorescence components, each electrophoresis tube is provided with a detection system involving an interference filer for selecting that of 340 to 390 nm from the fluorescence components. Outputs of such four detection systems are successively identified, to determine base sequences.

Base sequencing apparatus

An on-line system base sequencing apparatus wherein calibration coefficients for time bases of respective electrophoresis lanes are evaluated from differences between positions of signals already outputted in a range causing no sequence inversion and positions of substantially regular intervals for originally outputting signals, and time bases as to the respective electrophoresis lanes are calibrated with the calibration coefficients, thereby obtaining correct base sequence. Thus, the bases can be correctly sequenced even if electrophoresis speed differences are caused between the electrophoresis lanes.

With respect to a gel which is mounted on a gel electrophoresis apparatus, a calibration process is provided before starting of measurement to obtain calibration data for bringing a portion of the gel irradiated with excitation light onto the optical axis of a light receiving optical system for each scanning point, to perform measurement while controlling relative positional relation between the optical path of the excitation light and the optical axis of the light receiving optical system through the calibration data. Displacement, which may be inevitably caused between the optical axis of the light receiving optical system and the position irradiated with the excitation light by errors in working, assembling and adjustment, is basically eliminated, and sensitiveness irregularity is also removed.

Fluorescence detection type gel electrophoresis apparatus

An apparatus for determining a base sequence comprising an assembly for making an exciting beam incident on a slab of electrophoretic gel in a direction normal to the plane thereof, the gel containing nucleic acid fragments labeled with a labeling pigment and developed or being developed therein by electrophoresis, and a light receiving unit for receiving at an end face of the gel the luminescent emitted by the labeling pigment on the fragments developed in the gel.

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Kyoto, Japan

Hidehiko Fujii