KR20050006861A - The electrophoresis apparatus and the analysis system comprising the same - Google Patents

Abstract

PURPOSE: An electrophoresis system and an analyzing system containing the electrophoresis system are provided to observe the advancing situation of samples in a gel during the separated situation of the samples inside the electrophoresis system or an electrophoresis operation by producing the bottom of a container including the gel with a transparent material. CONSTITUTION: Buffering liquid is filled inside a container(20). A material separation is performed in a gel between electrodes inside the container. The bottom(21) of the container to include the gel is transparent to transmit ultraviolet rays. Thereby, an analysis is performed while observing an electrophoresis operation in a real time and a time from a preparation to a checking operation is reduced.

Description

ELECTROPHORESIS APPARATUS AND THE ANALYSIS SYSTEM COMPRISING THE SAME

The present invention relates to an electrophoretic device for separating and analyzing charged samples between electrodes and an analysis system including the same.

Electrophoresis refers to the movement of charged particles in opposite polarities in an electric field. The rate of movement is determined by several factors, including the amount of charge, the size and shape of the particles, the acidity (pH) and viscosity of the solution, the concentration and ionic strength of the other electrolytes in the solution, and the type of support. Thus, the rate of movement of charged particles in a solution depends on the nature of the molecule itself. Therefore, electrophoresis is used as a very effective means for separating or analyzing charged substances such as amino acids, nucleotides, proteins and the like.

Such electrophoretic devices are roughly classified into standard electrophoretic devices and gel electrophoretic devices.

The standard electrophoresis apparatus is configured to connect the buffers separated from each other with a gel tube as disclosed in Korean Patent No. 10-177013, and to immerse the electrodes in both buffers, respectively. This standard electrophoresis device has a problem that it is very difficult to classify DNA particles of different sizes.

Gel-type electrophoretic devices have been proposed to solve the above-mentioned problems, by placing a rod-shaped gel in a glass or plastic container ("gel container"), placing a sample on the top of the gel, and then applying an electric field. Consists of. The gel electrophoresis device is composed of a gel container and a cover, wherein two opposite electrodes are installed in parallel with each other at the lower ends of the gel container, and the gel is placed on the floor between the electrodes or on a gel bed disposed on the bottom. Put it up. At this time, a plurality of wells are formed on the upper surface of the gel in a direction perpendicular to the straight line connecting the electrodes.

Looking at the use of the gel-type electrophoresis device described above (Fig. 1), first, the agarose powder (2) and the buffer in the beaker (1) to boil, and when completely dissolved, poured into the gel caster (3) and hardens the gel. After filling the vessel of the electrophoretic device 4 with buffer and placing the sample in the gel well, a voltage is applied to both electrodes. The voltage is applied to the sample through the buffer, and the charged sample particles move through the buffer toward the electrode opposite to its charge. The moving distance and the moving speed of the sample at this time differ depending on the physical properties of the sample. For example, larger DNA molecules do not move away from the gel wells, while smaller ones move away from the wells of the gel to form DNA bands that vary in size. After the electrophoresis is completed, the gel is transferred to an UV Trans-Illuminator 5 and the ultraviolet rays are irradiated to the gel, and the DNA bands are visually observed by an image capture device (eg, a camera) and analyzed.

As described above, the existing sample (eg, DNA) analysis system is composed of an electrophoresis device, an ultraviolet light source (ultraviolet projector), an image capturing device (eg, a camera), and the like. The sample observation and analysis process via the image capture device 7 is performed at a separate location.

Since the container of the conventional electrophoretic device is made of a material that does not transmit ultraviolet rays, it was not possible to observe the running state of the sample on the gel in real time during electrophoresis through an ultraviolet lamp. Therefore, in order to solve the above problem, the present inventor manufactures the bottom of the container on which the gel is mounted with a transparent material which can transmit ultraviolet rays, so that the sample proceeds on the gel during the separation or electrophoresis of the sample in the electrophoresis device. The situation can be observed in real time, the present invention is based on this.

1 is a process flow diagram for analyzing DNA using a DNA analysis system including a conventional electrophoretic device, a UV Trans-Illuminator, and a camera.

Figure 2 is a perspective view showing a container of the electrophoretic device manufactured according to the present invention.

20: electrophoresis vessel 21: bottom of vessel on which gel is mounted

Figure 3 is a perspective view showing the installation position of the ultraviolet light source in the container of the electrophoretic device manufactured according to the present invention.

30: UV light source

Figure 4 is a perspective view of the cover of the electrophoretic device manufactured according to the present invention.

40: electrophoresis cover 41: cover portion corresponding to the portion on which the gel is to be mounted

5 is a perspective view showing a cover of the electrophoretic device manufactured according to the present invention.

50: electrophoresis cover 51: cover portion corresponding to the portion on which the gel is to be mounted

6A and 6B are perspective views of a camera hood made in accordance with the present invention.

60: camera hood 61: bottom of hood

7 is a view showing a gel caster according to the present invention.

7A is a perspective view of a gel placed on a gel caster, and FIG. 7B is a perspective view of a gel caster without a gel.

7C and 7D are sectional views of the gel caster from above, and FIG. 7E is a sectional view of the gel caster from the side.

70: gel caster

71: protrusion formed on the side of the gel caster

72: engraved groove formed on the side of the gel caster

73: hole formed in the side of the gel caster

74: negative groove formed in the bottom surface of the gel caster

75: hole formed in the bottom surface of the gel caster

80: gel

8 is a view showing an analysis system using an electrophoretic device manufactured according to the present invention.

The present invention provides an electrophoretic apparatus in which a buffer is filled in a container and material is separated on a gel between opposing electrodes in the container, wherein the bottom of the container on which the gel is mounted is made of a transparent material capable of transmitting ultraviolet rays. Provides electrophoresis device.

In addition, the present invention is made of a transparent material that can transmit light emitted from the sample to be analyzed, or only a portion corresponding to the upper end of the portion on which the gel is mounted in the container can transmit light emitted from the sample to be analyzed Provided is a container cover for an electrophoretic device made of a transparent material or perforated by the size of a gel to be mounted in the container.

Moreover, this invention provides the container cover which has a convex downward groove in the part corresponding to the upper end of the part in which the gel in a container is mounted.

In addition, the present invention provides a support of an image capture device having a plate made of a transparent material through which the bottom surface thereof can transmit light emitted from a sample to be analyzed.

Hereinafter, a gel type electrophoretic apparatus and a sample analysis system including the same according to a preferred embodiment of the present invention will be described in detail with reference to the accompanying drawings.

In general, in order to confirm the location of the sample containing DNA on the gel during or after electrophoresis, fluorescence, that is, a phenomenon in which the material emits light by stimulation of light is used.

Usually, Intercalate type (staining DNA into two chains of DNA) such as ethidium bromide (ET-BR) is used for DNA and RNA staining. Ultraviolet rays are used for color development.

The staining time can be stained by 1) ET-BR directly in the sample, 2) ET-BR in the gel, or 3) electrophoresis and then gel in the ET-BR solution. ET-BR is often used inside.

As shown in FIG. 2, the electrophoretic apparatus of the present invention is characterized in that the bottom of the container 21 on which the gel is mounted is made of a transparent material through which ultraviolet rays can be transmitted.

Thereby, an ultraviolet light source such as an ultraviolet lamp or an ultraviolet projector is installed under the container of the electrophoretic device, thereby irradiating ultraviolet rays onto the gel through the bottom of the container, so that the path of the electrophoretic or electrophoretic gel phase sample is carried out in real time. Speed and separation can be checked and analyzed.

For example, an ultraviolet ray projector is equipped with an ultraviolet light tube (UV tube) and a UV filter that transmits only ultraviolet light and absorbs all visible light on its glass surface. Therefore, an electrophoretic tank (Tank) can be made on the UV projector, and an UV projector directly attached to the electrophoretic device can be made.

On the other hand, it is preferable that the base 21 of the container on which the gel is mounted has a concave-convex shape that is recessed (FIG. 2). Corresponding to the portion where the bottom 11 of the container on which the gel is mounted is recessed, an ultraviolet lamp 21 that is an ultraviolet source that can be inserted into the recess of the bottom of the container can be used (see Fig. 3).

In addition, the electrophoretic apparatus of the present invention is made of a transparent material that can transmit light emitted from a sample to be analyzed (eg, ultraviolet light, visible light, infrared light), or a portion corresponding to an upper end of a portion on which a gel is mounted in a container. It is possible to use a container cover made of a transparent material through which light emitted from the sample to be analyzed can be transmitted (41) or penetrated (41) by the size of a gel mounted in the container.

At this time, it is preferable that the upper plate of the container cover has a convex downward groove 51 at a portion corresponding to the upper end of the portion where the gel in the container is mounted (FIG. 5). When observing the separation of the sample in the vessel of the electrophoresis device, the more buffer, the greater the scattering of light, which impedes the identification of bands (eg, DNA bands) of the separated sample. Therefore, as in the present invention, the container cover has a convex downward groove 51 at a portion corresponding to the upper end of the portion in which the gel is mounted in the container, and the groove portion emits light emitted from a sample to be analyzed (eg, ultraviolet light, visible light). With a plate made of a transmissive material capable of transmitting light, infrared rays), the buffer layer positioned between the image capturing device and the gel by placing the convex groove portion in the electrophoretic buffer and placed on the gel By reducing the thickness of the light scattering by the buffer can be reduced.

Non-limiting examples of the transparent material that can transmit ultraviolet rays include PS (polystyrene), ABS (acrylonitrile-butadiene-styrene resin), PETE, PC (polycarbonate), acrylic type (PMMA (polymethylmethacryl) Rate), MMA (methyl methacrylate)), transparent PP (polypropylene), transparent PE (polyethylene), glass, and the like.

In the present invention, when checking or analyzing a state of a sample separated by electrophoresis using an ultraviolet light source, the bottom surface of the sample may transmit light emitted from the sample to be analyzed (eg, ultraviolet light, visible light, infrared light). It is preferable to use a support (eg camera hood) of an image capture device having a plate 61 made of a transparent material that can be used (FIG. 6). In order to solve the problem of light scattering by the buffer solution, if the bottom of the support of the image capturing device (i.e., the electrophoretic device) is provided with a plate made of a transmissive material that can transmit ultraviolet rays, By soaking the underside in an electrophoretic buffer and placing it on the gel, it is possible to reduce the scattering of light by the buffer by reducing the thickness of the buffer layer located between the image capture device and the gel.

Image capturing devices include cameras, camcorders, digital cameras, and the like. On the other hand, it is also possible to use an underwater camera without the container cover of the electrophoretic device.

On the other hand, when the gel used for electrophoresis is produced in a gel caster, a protruding portion on the side or bottom inside the gel caster adjacent to the gel to prevent separation of the gel caster and the gel during electrophoresis or movement of the gel caster. It is desirable to make 71 or holes 73 and 75 or recessed grooves 72 and 74. If the gel is to be electrophoresed on the gel caster, the gel caster is also made of a transparent material through which ultraviolet light can be transmitted.

While the present invention has been described with reference to the preferred embodiments, those skilled in the art will understand that the present invention may be modified without departing from the spirit and scope of the invention.

By using the electrophoretic apparatus of the present invention, it is possible to analyze while observing the electrophoretic operation in real time, it is possible to shorten the time required for the preparation and confirmation process from the existing electrophoretic operation to the confirmation process, and according to a long time operation It can reduce the possibility of error. It also enables automated DNA analysis.

Claims (10)

An electrophoretic device in which a buffer is filled in a container and material is separated on a gel between opposing electrodes in the container, wherein the bottom of the container on which the gel is mounted is made of a transparent material capable of transmitting ultraviolet rays. The electrophoretic apparatus according to claim 1, wherein the bottom of the container on which the gel is mounted has a concave-convex shape that is recessed. The electrophoretic apparatus according to claim 1, wherein an ultraviolet light source is attached to the bottom of the bottom of the container on which the gel is mounted. 3. An electrophoretic apparatus according to claim 2, wherein an ultraviolet light source is inserted into a portion where the bottom of the container on which the gel is mounted is recessed. The method of claim 1, wherein the light emitted from the sample to be analyzed may be transmitted, or only a portion corresponding to the upper end of the portion in which the gel is mounted in the container may be transmitted. Electrophoretic device further comprises a container cover made of a transparent material, or perforated by the size of a gel mounted in the container. 6. The electrophoretic apparatus according to claim 5, wherein the container cover has a convex downward groove at a portion corresponding to an upper end of the portion in which the gel is mounted in the container, and a portion of the groove may be contained in the buffer in the container. The transparent material that can transmit ultraviolet rays is PS (polystyrene), ABS (acrylonitrile-butadiene-styrene resin), PETE, PC (polycarbonate), PMMA (polymethylmethacrylate) , Electrophoretic device characterized in that selected from the group consisting of MMA (methyl methacrylate), transparent PP (polypropylene), transparent PE (polyethylene), and glass. A support of an image capture device, the bottom of which has a plate made of a transparent material that can transmit light emitted from a sample to be analyzed. A gel caster having protrusions or holes or recessed grooves on the side or bottom of the gel caster adjacent to the gel. 10. The gel caster of claim 9 made of a transparent material capable of transmitting ultraviolet light.