Polynucleotide probe chip and polynucleotide detection method

A method for detecting polynucleotides comprising the steps of:

• labeling sample polynucleotides with fluorophores;
• adding the fluorophore-labeled sample polynucleotides to a polynucleotide probe chip comprising plural areas having gels immobilizing different polynucleotide probes, and comprising first and second electrodes for migrating the fluorophore-labeled sample polynucleotides in the gels, between which the plural areas are arranged;
• changing, plural times, polarities of the first and second electrodes, and migrating the fluorophore-labeled sample polynucleotides in the gels of the respective areas by electrophoresis, to hybridize the polynucleotide probes immobilized in the gels with the fluorophore-labeled sample polynucleotides and to form hybrids;
• and
• detecting the fluorophore-labeled sample polynucleotides forming the hybrids in the respective areas.

A method for detecting polynucleotides comprising the steps of:

• adding sample polynucleotides to a polynucleotide probe chip comprising plural areas having gels immobilizing different polynucleotide probes, and comprising first and second electrodes for migrating the sample polynucleotides in the gels, between which the plural area are arranged;
• changing, plural times, polarities of the first and second electrodes, and migrating the sample polynucleotides in the gels of the respective areas by electrophoresis, to hybridize the polynucleotide probes immobilized in the gels with the sample polynucleotides and to form hybrids;
• introducing, by using DNA polymerase, a fluorophore-labeled dNTP or a fluorophore-labeled ddNTP into extended chains of the polynucleotide probes forming the hybrids to form fluorophore-labeled polynucleotide probes;
• and
• detecting the fluorophore-labeled polynucleotide probes forming the hybrids in the respective areas.

A polynucleotide detecting device comprising:

• a polynucleotide probe chip comprising an optically transparent substrate having a concave portion;
• gels immobilized in holes which are formed in a flat surface of the concave portion, penetrate the substrate and have tapered side faces;
• different polynucleotide probes immobilized in the gels;
• and a portion for irradiating a side surface of the substrate with laser light;
• an electrode vessel in which a first electrode is located, and in which is a lower surface of the polynucleotide probe chip;
• a second electrode which is set in a solution including sample polynucleotides added onto a surface of the polynucleotide probe chip;
• and
• a switch which changes, plural times, polarities of the first and second electrodes, to migrate the sample polynucleotides in the gels of the respective holes by electrophoresis, thereby hybridizing the polynucleotide probes immobilized in the gels with the sample polynucleotides;
• a means for irradiating the gels of the respective holes with laser light, along a direction parallel to a main surface of the substrate;
• and
• light detector for detecting emitted fluorescence emitted in the respective holes form a direction perpendicular to the main surface of the substrate.

A method for detecting polynucleotides using a polynucleotide probe chip comprising plural areas having gels immobilizing different polynucleotide probes, and comprising first and second electrodes for migrating sample polynucleotides in the gels, between which the plural areas are arranged, the method comprising the steps of:

• changing, plural times, polarities of the first and second electrodes, and migrating the sample polynucleotides in the gels of the respective areas by electrophoresis, thereby hybridizing the polynucleotide probes immobilized in the gels with the sample polynucleotides;
• irradiating the gels of the areas simultaneously with laser light, along a direction parallel to a main surface of a substrate of the polynucleotide probe chip;
• and
• detecting emitted fluorescence along a direction perpendicular to the main surface of the substrate.

A polynucleotide detecting device comprising:

• a polynucleotide probe chip comprising plural areas having gels immobilizing different polynucleotide probes, on surfaces of respective areas of a substrate of the polynucleotide probe chip;
• photodiodes formed on surfaces of the respective areas;
• and first and second electrodes for migrating sample polynucleotides in the gels, between which the plural areas are arranged;
• a switch which changes, plural times, polarities of the first and second electrodes, to migrate the sample polynucleotides in the gels of the respective areas by electrophoresis, thereby hybridizing the polynucleotide probes immobilized in the gels with the sample polynucleotides;
• and
• a means for irradiating the gels of the areas with laser light, along a direction parallel to a main surface of the substrate.

A polynucleotide probe chip comprising:

• a substrate having a concave portion;
• plural portions which are arranged in one direction and immobilize gels in a flat base face the concave portion:
• first and second electrodes between which the plural portions are arranged;
• different polynucleotide probes immobilized in the gels;
• a portion for irradiating a side surface of the substrate with laser light;
• and
• first and second electrodes for migrating sample polynucleotides in the gels, between which the plural portions are arranged, polarities of the first and second electrodes being changed, plural times, to migrate the sample polynucleotides in the gels of the respective potions by electrophoresis, thereby hybridizing the polynucleotide probes immobilized in the gels with the sample polynucleotides.

A polynucleotide probe chip comprising:

• plural areas having gels immobilizing different polynucleotide probes, on surfaces of respective areas of a substrate of the polynucleotide probe chip;
• photodiodes formed on surfaces of the respective areas;
• band pass filters formed over the respective photodiodes;
• first electrodes formed over the respective band pass filters;
• and
• second electrodes between which the plural areas are arranged, polarities of the first and second electrodes being changed, plural times, to migrate sample polynucleotides in the gels of the respective areas by electrophoresis, thereby hybridizing the polynucleotide probes immobilized in the gel(s) with the sample polynucleotides.

A polynucleotide detecting device comprising:

• a polynucleotide probe chip comprising plural areas having gels immobilizing different polynucleotide probes on surfaces of respective areas of a substrate of the polynucleotide probe chip;
• photodiodes formed on surfaces of the respective areas;
• first electrodes formed over the respective band pass filters, between which insulating layers are arranged;
• and
• means for changing, plural times, polarities of the first and second electrodes to migrate sample polynucleotides in the gels of the respective areas by electrophoresis, thereby hybridizing the polynucleotide probes immobilized in the gels with the sample polynucleotides;
• and
• a means for irradiating the gels of the areas with laser light, along a direction parallel to a main surface of the substrate.

A polynucleotide probe chip comprising:

• plural areas having gels immobilizing different polynucleotide probes and being arranged in one direction;
• first electrodes arranged in the respective areas;
• and
• second electrodes between which the plural areas are arranged in the one direction;
• wherein a high frequency electrical potential is applied between the first and second electrodes and migrating sample polynucleotides migrate in the gels of the respective areas by electrophoresis, to hybridize the polynucleotide probes immobilized in the gels with the sample polynucleotides.

A polynucleotide probe chip comprising:

• plural areas having gels immobilizing different polynucleotide probes and being arranged in one direction;
• first electrodes arranged in the respective areas;
• second electrodes between which the plural areas are arranged in the one direction;
• and
• third electrodes arranged between the first and second electrodes;
• wherein a high frequency electrical potential is applied between the first and second electrodes and sample polynucleotides migrate in the gels of the respective areas by electrophoresis, to hybridize the polynucleotide probes immobilized in the gels with the sample polynucleotides, and, in addition, a high frequency electrical potential is applied between the second and third electrodes to migrate, by electrophoresis, the sample polynucleotides which are not hybridized with the polynucleotide probes immobilized in the gels from the gels.

A polynucleotide detecting device comprising:

• a polynucleotide probe chip comprising plural areas having gels immobilizing different polynucleotide probes, and comprising first and second electrodes for migrating sample polynucleotides in the gels, between which the plural area are arranged;
• a means for changing, plural times, polarities of the first and second electrodes to migrate the sample polynucleotides in the gels of the respective areas by electrophoresis, thereby hybridizing the polynucleotide probes immobilized in the gels with the sample polynucleotides;
• a means for releasing the sample polynucleotides from the polynucleotide probes with which the sample polynucleotides are hybridized;
• and
• capillaries for separating the released sample polynucleotides by electrophoresis, ends of the capillaries being arranged at intervals same as intervals of the respective areas.

A method detecting polynucleotides comprising the steps of:

• adding sample polynucleotides to a polynucleotide probe chip comprising plural areas having gels immobilizing different polynucleotide probes and being arranged in one direction;
• first electrodes arranged in the respective areas;
• and second electrodes between which the plural areas are arranged in the one direction;
• and
• applying a high frequency electrical potential between the first and second electrodes and migrating the sample polynucleotides in the gels of the respective areas by electrophoresis, thereby hybridizing the polynucleotide probes immobilized in the gels with the sample polynucleotides.

A polynucleotides assay method comprising the steps of:

• adding sample polynucleotides on a surface of a polynucleotide probe chip comprising plural areas having gels immobilizing different polynucleotide probes and being arranged in a one direction;
• first electrodes arranged in the respective areas;
• and second electrodes between which the plural areas are arranged in the one direction;
• and third electrodes arranged between the first and second electrodes;
• applying a high frequency electrical potential between the first and second electrodes and migrating the sample polynucleotides in the gels of the respective areas by electrophoresis, to hybridize the polynucleotide probes immobilized in the gels with the sample polynucleotides;
• and
• applying a high frequency electrical potential between the second and third electrodes to migrate, by electrophoresis, the sample polynucleotides which are not hybridized with the polynucleotide probes immobilized in the gels from the gels.

A polynucleotide assay method comprising the steps of:

• adding sample polynucleotides to a polynucleotide probe chip comprising plural areas having gels immobilizing different polynucleotide probes, and comprising first and second electrodes for migrating the sample polynucleotides in the gels, between which the plural areas are arranged;
• changing, plural times, polarities of the first and second electrodes to migrate the sample polynucleotides in the gels of the respective areas by electrophoresis, thereby hybridizing the polynucleotide probes immobilized in the gels with the sample polynucleotides;
• releasing the sample polynucleotides from the polynucleotide probes with which the sample polynucleotides are hybridized;
• and
• using capillaries arranged at intervals same as intervals of the respective areas, collecting and separating the released sample polynucleotides by electrophoresis.

A polynucleotide probe chip comprising:

• plural areas having gels immobilizing different polynucleotide probes, the plural areas being arranged in one direction;
• first and second electrodes for migrating sample polynucleotides in the gels, between which the plural areas are arranged, the first and second electrodes being arranged in the one direction;
• and
• photodiodes formed on surfaces of the respective areas.

A polynucleotide detecting device comprising:

• a polynucleotide probe chip comprising plural areas having gels immobilizing different polynucleotide probes, the plural areas being arranged in one direction;
• photodiodes formed on surfaces of the respective areas;
• and first and second electrodes for migrating sample polynucleotides in the gels, between which the plural areas are arranged, the first and second electrodes being arranged in the one direction;
• and
• a means for irradiating the gels of the areas with laser light, along a direction parallel to a main surface of a substrate of the polynucleotide probe chip.

A polynucleotide probe chip comprising:

• plural areas having gels immobilizing different polynucleotide probes, the plural areas being arranged in one direction;
• first and second electrodes for migrating sample polynucleotides in the gels, between which the plural areas are arranged, the first and second electrodes being arranged in the one direction;
• photodiodes formed on surfaces of the respective areas;
• and
• band pass filters formed on surfaces of the respective photodiodes through respective insulating layers.

A polynucleotide detecting device comprising:

• a polynucleotide probe chip comprising plural areas having gels immobilizing different polynucleotide probes, the plural areas being arranged in one direction;
• first and second electrodes for migrating sample polynucleotides in the gels, between which the plural areas are arranged, the first and second electrodes being arranged in the one direction;
• photodiodes formed on surfaces of the respective areas;
• and band pass filters formed on surfaces of the respective photodiodes through respective insulating layers;
• a means for irradiating the gels of the areas with laser light, along a direction parallel to a main surface of a substrate of the polynucleotide probe chip.

A polynucleotide probe chip comprising:

• plural areas having gels immobilizing different polynucleotide probes, the plural areas being arranged in one direction;
• first and second electrodes between which the plural areas are arranged, the first and second electrodes being arranged in the one direction;
• photodiodes formed on surfaces of the respective areas;
• band pass filters formed on surfaces of the respective photodiodes through respective insulating layers;
• and
• transparent electrodes formed on the band pass filters through respective insulating layers.

A polynucleotide detecting device comprising:

• a polynucleotide probe chip comprising plural areas having gels immobilizing different polynucleotide probes, the plural areas being arranged in one direction;
• first and second electrodes between which the plural areas are arranged, the first and second electrodes being arranged in the one direction;
• photodiodes formed on surfaces of the respective areas;
• band pass filters formed on surfaces of the respective photodiodes through respective insulating layers;
• transparent electrodes formed on the band pass filters through respective insulating layers;
• and
• a means for irradiating the gels of the areas with laser light, along a direction parallel to a main surface of a substrate of the polynucleotide probe chip.

A polynucleotide probe chip comprising:

• plural areas having gels immobilizing different polynucleotide probes, the plural areas being arranged in one direction;
• first and second electrodes between which the plural areas are arranged, the first and second electrodes being arranged in the one direction;
• first point electrodes formed on surfaces of the respective areas;
• second point electrodes formed on a surface between the first point electrode and the first electrode;
• and
• third point electrodes formed on a surface between the first point electrode and the second point electrode.

A polynucleotide detecting device comprising:

• a polynucleotide probe chip comprising plural areas having gels immobilizing different polynucleotide probes, the plural areas being arranged in one direction;
• first and second electrodes between which the plural areas are arranged, the first and second electrodes being arranged in the one direction;
• first point electrodes formed on surfaces of the respective areas;
• second point electrodes formed on a surface between the first point electrode and the first electrode;
• third point electrodes formed on a surface between the first point electrode and the second electrode;
• a means for irradiating the gels of the respective areas with laser light, along a direction parallel to a main surface of a substrate of the polynucleotide probe chip;
• and
• light detector for detecting emitted fluorescence emitted in the respective areas form a direction perpendicular to the main surface of a substrate of the polynucleotide probe chip.