Apparatus for the separation and fractionation of differentially expressed gene fragments

An apparatus for the separation and fractionation of differentially expressed gene fragments, the apparatus comprising:

• a separating means including a capillary filled with a separation medium to separate DNA fragments by electrophoresis, the DNA fragments each being labeled with a fluorophore;
• a detecting means to apply laser light to the DNA fragments separated in the capillary and to detect fluorescence emitted from the fluorophore;
• a means to form a sheath flow of a buffer solution to carry the separated DNA fragments eluted from the end of the capillary;
• a sampling means including sampling vessels to fractionate and sample the separated DNA fragments, according to their size;
• a transferring means to transfer the buffer solution containing the separated DNA fragments to the sampling means, the transferring means including a sampling tube which is disposed such that its first end opposes the end of the capillary at a specified gap, the separated DNA fragments being transferred to the opening of the first end of the sampling tube;
• a reservoir, having disposed therein a second end of the sampling tube, to contain the buffer solution eluted from the second end of the sampling tube, the reservoir having, at its bottom, an aperture having such a diameter that the sampling tube can be placed therethrough;
• a first tube to transport a washing fluid to the reservoir;
• a second tube to drain the buffer solution from the reservoir;
• and
• a control means to control the sampling means based on a signal gained by the detecting means;
• wherein the second end of the sampling tube is placed through the aperture to transport the separated DNA fragments transferred by the sampling tube to the sampling vessels;
• and
• wherein a voltage for the electrophoresis and a length of the capillary are adjusted such that a spread in time of the separated DNA fragments caused by the transferring means during the transfer of the separated DNA fragments to the sampling vessels is smaller than a difference in separation time of the DNA fragments in the separating means.

An apparatus for the separation and fractionation of differentially expressed gene fragments, the apparatus comprising:

• a separating means including a plurality of capillaries each filled with a separation medium to separate DNA fragments by electrophoresis, the DNA fragments each being labeled with a fluorophore;
• a voltage applying means to apply a voltage for electrophoresis to each of the capillaries independently;
• a detecting means to apply laser light to the DNA fragments separated by each of the capillaries and to detect fluorescence emitted from the fluorophore;
• a means to form a sheath flow of a buffer solution to carry the separated DNA fragments eluted from the end of each of the capillaries;
• a sampling means including sampling vessels to fractionate and sample the separated DNA fragments, according to their size;
• a transferring means to transfer the buffer solution containing the sampled DNA fragments to the sampling means, the transferring means including a sampling tube which is disposed such that its end opposes the end of each of the capillaries at a specified gap, the separated DNA fragments being transferred to the opening of the end of the sampling tube;
• and
• a control means to control the sampling means based on a signal gained by the detecting means;
• wherein the voltage applying means changes the voltage during electrophoresis based on the signal gained by the detecting means;
• and
• wherein the voltage for the electrophoresis and a length of each of the capillaries are adjusted such that a spread in time of the separated DNA fragments caused by the transferring means during the transfer of the separated DNA fragments to the sampling vessels is smaller than a difference in separation time of the DNA fragments in the separating means.

An apparatus for the separation and fractionation of differentially expressed gene fragments, said apparatus comprising:

• a separating means including a capillary filled with a separation medium to separate DNA fragments by electrophoresis, said DNA fragments each being labeled with a fluorophore,
• a detecting means to apply laser light to the DNA fragments separated by the capillary and to detect fluorescence emitted from the fluorophore,
• a transferring means including a sampling tube which is placed with its first end opposed to the terminal end of the electrophoresis capillary at a specified gap, said separated DNA fragments being transferred to the opening of said first end,
• a means to form a sheath flow of a buffer solution to carry the separated DNA fragments eluted from the capillary to the opening of said first end,
• a sampling means having sampling vessels to fractionate and sample said DNA fragments according to their sizes, said DNA fragments transferred by the sampling tube, and
• a control means to control the sampling means based on a signal gained by the detecting means, wherein
• said sampling tube has an inner diameter greater than the outer diameter of said capillary.

An apparatus for the separation and fractionation of differentially expressed gene fragments, comprising:

• a separating means including a capillary filled with a separation medium to separate DNA fragments by electrophoresis, said DNA fragments each labeled with a fluorophore,
• a detecting means to apply laser light to the separated DNA fragments by the capillaries and to detect fluorescence emitted from the fluorophore,
• a first tray to hold wells for holding a sampling solution containing said DNA fragments,
• a second tray to hold wells for holding a marker solution each containing a marker,
• an injecting means to inject said sample solution and said marker solution into said capillary separately,
• a transferring means including a sampling tube which is placed with its first end opposed to the end of the electrophoresis capillary at a specified gap, said separated DNA fragments being transferred to the opening of said first end,
• a means to form a sheath flow of a buffer solution to carry the separated DNA fragments eluted from said capillary to the opening of the first end,
• a sampling means including sampling vessels to fractionate and sample the DNA fragments transferred by the sampling tube according to their sizes, and
• a control means to control said sampling means based on a signal gained by said detecting means, wherein the sampling tube has a length ranging from 5 cm to 15 cm and an inner diameter ranging from 50 μm to 100 μm, and wherein said buffer solution flowing through the sampling tube has a flow rate of about 10 mm/sec.

An apparatus for the separation and fractionation of differentially expressed gene fragments, the apparatus comprising:

• a capillary which is filled with a separation medium to separate by electrophoresis DNA fragments each being labeled with a fluorophore;
• a laser light source which generates a laser light to apply to the DNA fragments separated by electrophoresis in the capillary;
• a two-dimensional detector which detects fluorescence emitted from the fluorophore;
• transferring means including a sampling tube which is disposed such that an end of the sampling tube opposes an end of the capillary at a specified gap, separated DNA fragments eluted from the end of the capillary being transferred to an opening in the end of the sampling tube;
• means for forming a sheath flow of a buffer solution to carry the separated DNA fragments eluted from the end of the capillary to the opening in the end of the sampling tube;
• sampling means including sampling vessels to fractionate and sample the separated DNA fragments, according to their size, the separated DNA fragments being transferred by the sampling tube;
• and
• control means for controlling the sampling means based on the fluorescence detected by the two-dimensional detector;
• wherein the transferring means further includes
• means for forming droplets of the buffer solution containing the DNA fragments separated in the capillary, and
• transporting means for transporting the formed droplets to the sampling vessels by gas flow.

An apparatus for the separation and fractionation of differentially expressed gene fragments, the apparatus comprising:

• separating means including a capillary filled with a separation medium to separate DNA fragments by electrophoresis, the DNA fragments each being labeled with a fluorophore;
• detecting means for applying laser light to the DNA fragments separated in the capillary and for detecting fluorescence emitted from the fluorophore;
• transferring means including a sampling tube which is disposed such that a first end of the sampling tube opposes an end of the capillary at a specified gap, separated DNA fragments eluted from the end of the capillary being transferred to an opening in the first end of the sampling tube;
• means for forming a sheath flow of a buffer solution to carry the separated DNA fragments eluted from the end of the capillary to the opening in the first end of the sampling tube;
• sampling means including sampling vessels to fractionate and sample the separated DNA fragments, according to their size, the separated DNA fragments being transferred by the sampling tube;
• and
• a reservoir, having disposed therein a second end of the sampling tube, to contain buffer solution eluted from the second end of the sampling tube, a bottom of the reservoir having an aperture having a diameter enabling the sampling tube to be placed therethrough;
• a first tube to transport a washing fluid to the reservoir;
• a second tube to drain the buffer solution from the reservoir;
• and
• control means for controlling the sampling means based on a signal produced by the detecting means;
• wherein the second end of the sampling tube is placed through the aperture to transport the separated DNA fragments transferred by the sampling tube to the sampling vessels.

An apparatus for the separation and fractionation of differentially expressed gene fragments, the apparatus comprising:

• separating means including a capillary filled with a separation medium to separate DNA fragments by electrophoresis, the DNA fragments each being labeled with a fluorophore;
• detecting means for applying laser light to the DNA fragments separated in the capillary and for detecting fluorescence emitted from the fluorophore;
• transferring means including a sampling tube which is disposed such that an end of the sampling tube opposes an end of the capillary at a specified gap, separated DNA fragments eluted from the end of the capillary being transferred to an opening in the end of the sampling tube;
• means for forming a sheath flow of a buffer solution to carry the separated DNA fragments eluted from the end of the capillary to an opening in the end of the sampling tube;
• sampling means including sampling vessels to fractionate and sample the separated DNA fragments, according to their size, the separated DNA fragments being transferred by the sampling tube;
• and
• control means for controlling the sampling means based on a signal produced by the detecting means;
• wherein the laser light is applied to the capillary.

An apparatus for the separation and fractionation of differentially expressed gene fragments, the apparatus comprising:

• separating means including a plurality of capillaries each filled with a separation medium to separate DNA fragments by electrophoresis, the DNA fragments each being labeled with a fluorophore;
• voltage applying means for applying a voltage for electrophoresis to each of the capillaries independently;
• detecting means for applying laser light to the DNA fragments separated in each of the capillaries and for detecting fluorescence emitted from the fluorophore;
• transferring means including a sampling tube which is disposed such that an end of the sampling tube opposes an end of each of the capillaries at a specified gap, separated DNA fragments eluted from the end of each of the capillaries being transferred to an opening in the end of the sampling tube;
• means for forming a sheath flow of a buffer solution to carry the separated DNA fragments eluted from the end of each of the capillaries to the opening in the end of the sampling tube;
• sampling means including sampling vessels to fractionate and sample the separated DNA fragments, according to their size, the separated DNA fragments being transferred by the sampling tube;
• and
• control means for controlling the sampling means based on a signal produced by the detecting means;
• wherein the voltage applying means changes the voltage during electrophoresis based on the signal produced by the detecting means.

An apparatus for the separation and fractionation of differentially expressed gene fragments, the apparatus comprising:

• separating means including a capillary filled with a separation medium to separate DNA fragments by electrophoresis, the DNA fragments each being labeled with a fluorophore;
• detecting means for applying laser light to the DNA fragments separated in the capillary and for detecting fluorescence emitted from the fluorophore;
• transferring means including a sampling tube which is disposed such that an end of the sampling tube opposes an end of the capillary at a specified gap, separated DNA fragments eluted from the end of the capillary being transferred to an opening in the end of the sampling tube;
• means for forming a sheath flow of a buffer solution to carry the separated DNA fragments eluted from the end of the capillary to the opening in the end of the sampling tube;
• sampling means including sampling vessels to fractionate and sample the separated DNA fragments, according to their size, the separated DNA fragments being transferred by the sampling tube;
• and
• control means for controlling the sampling means based on a signal produced by the detecting means;
• wherein the sampling tube has a length ranging from 5 cm to 15 cm and an inner diameter ranging from 50 μm to 100 μm;
• and
• wherein buffer solution flowing through the sampling tube has a flow rate about 10 mm/sec.

An apparatus for the separation and fractionation of differentially expressed gene fragments, the apparatus comprising:

• a capillary which is filled with a separation medium to separate by electrophoresis DNA fragments each being labeled with a fluorophore;
• a laser light source which generates a laser light to apply to the DNA fragments separated by electrophoresis in the capillary;
• a two-dimensional detector which detects fluorescence emitted from the fluorophore;
• a sampling tube which is disposed such that an end of the sampling tube opposes an end of the capillary at a specified gap, separated DNA fragments eluted from the end of the capillary being carried through a sheath flow of a buffer solution into the sampling tube to be transferred by the sampling tube;
• a fraction collector which has sampling vessels to fractionate and sample the separated DNA fragments transferred by the sampling tube, according to their size;
• and
• a controller which controls a movement of the fraction collector based on the fluorescence detected by the two-dimensional detector;
• wherein the sampling tube has an inner diameter greater than an outer diameter of the capillary.

An apparatus for the separation and fractionation of differentially expressed gene fragments, the apparatus comprising:

• a capillary which is filled with a separation medium to separate by electrophoresis DNA fragments each being labeled with a fluorophore;
• a laser light source which generates a laser light to apply to the DNA fragments separated by electrophoresis in the capillary;
• a two-dimensional detector which detects fluorescence emitted from the fluorophore;
• a sampling tube which is disposed such that an end of the sampling tube opposes an end of the capillary at a specified gap, separated DNA fragments eluted from the end of the capillary being carried through a sheath flow of a buffer solution into the sampling tube to be transferred by the sampling tube;
• a fraction collector which has sampling vessels to fractionate and sample the separated DNA fragments transferred by the sampling tube, according to their size;
• and
• a controller which controls a movement of the fraction collector based on the fluorescence detected by the two-dimensional detector;
• wherein the laser light is applied to the capillary.

An apparatus for the separation and fractionation of differentially expressed gene fragments, the apparatus comprising:

• a capillary which is filled with a separation medium to separate by electrophoresis DNA fragments each being labeled with a fluorophore;
• a laser light source which generates a laser light to apply to the DNA fragments separated by electrophoresis in the capillary;
• a two-dimensional detector which detects fluorescence emitted from the fluorophore;
• a sampling tube which is disposed such that a first end of the sampling tube opposes an end of the capillary at a specified gap, separated DNA fragments eluted from the end of the capillary being carried through a sheath flow of a buffer solution into an opening in the first end of the sampling tube to be transferred by the sampling tube;
• a reservoir, having disposed therein a second end of the sampling tube, to contain buffer solution eluted from the second end of the sampling tube, a bottom of the reservoir having an aperture having a diameter enabling the sampling tube to be placed therethrough;
• a first tube to transport a washing fluid to the reservoir;
• a second tube to drain the buffer solution from the reservoir;
• a fraction collector which has sampling vessels to fractionate and sample the separated DNA fragments transferred from an opening in the second end of the sampling tube, according to their size;
• and
• a controller which controls a movement of the fraction collector based on the fluorescence detected by the two-dimensional detector;
• wherein the second end of the sampling tube is placed through the aperture to transport the separated DNA fragments transferred by the sampling tube to the sampling vessels.

An apparatus for the separation and fractionation of differentially expressed gene fragments, the apparatus comprising:

• a capillary which is filled with a separation medium to separate by electrophoresis DNA fragments each being labeled with a fluorophore;
• a laser light source which generates a laser light to apply to the DNA fragments separated by electrophoresis in the capillary;
• a two-dimensional detector which detects fluorescence emitted from the fluorophore;
• a sampling tube which is disposed such that a first end of the sampling tube opposes an end of the capillary at a specified gap, separated DNA fragments eluted from the end of the capillary being carried through a sheath flow of a buffer solution into an opening in the first end of the sampling tube to be transferred by the sampling tube;
• a fraction collector which has sampling vessels to fractionate and sample the separated DNA fragments transferred from an opening in a second end of the sampling tube, according to their size;
• a conical shaped member which has a hollow space through which the second end of the sampling tube passes;
• and
• a controller which controls a movement of the fraction collector based on the fluorescence detected by the two-dimensional detector;
• wherein droplets of the buffer solution containing the separated DNA fragments are formed by flowing a gas into the hollow space;
• and
• wherein the formed droplets are transported to the sampling vessels by gas flow.