JPH0526845A - Gel electrophoretic cell for detecting light - Google Patents

Abstract

PURPOSE:To obtain a gel electrophoretic cell for the analysis of DNA or the like using fluorescence detection characterized in that the forming labor of a gel is reduced to a large extent, fluorescence can be stably detected with high sensitivity and a wide effective using area can be taken on a migration plate. CONSTITUTION:A gel electrophoretic cell for detection is equipped with at least two buffer liquid tanks 1 and gell migration plates 2, 3 constituted of the gel 4 held between glass plates. The side end surface parts of the gel migration plates 2, 3 are covered with a part of the buffer liquid tanks 1.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a gel electrophoresis tank for photodetection.
(Hereinafter, also simply referred to as "electrophoresis tank"), especially DNA, R
The present invention relates to a migration tank for a real-time fluorescence detection electrophoretic device for NA or protein.

[0002]

2. Description of the Related Art Conventionally, for the separation and detection of DNA and the like, DNA is labeled with a radioactive isotope and separated by gel electrophoresis.
This was done by transferring the separation pattern to a film (autoradiography). However, in this system or device, a device for labeling DNA or the like with fluorescence and performing electrophoresis / separation for real-time detection has been developed in recent years due to the complexity of using radioactive isotopes and the difficulty of taking time. (For example, “Opti
mization of Parameters in a DNA Seqenator Usi
ng Fluorescence Detection ”(Bio / Tech 6 , 816-82
1, 1988)). Among the several reported devices, the device that uses a laser for excitation of fluorescence from the side of the separation gel can irradiate a long region, that is, many migration paths at the same time, so the average excitation light per migration path is There is an advantage that the irradiation amount is increased and high sensitivity is obtained. In the migration plate used here, a glass window for laser incidence was usually attached to a side light incidence part to prepare a gel for use. In addition, gel is made by separating two glass plates or quartz plates with spacers of about 0.3 mm thickness, sealing both sides and bottom with adhesive tape, and pouring the gel material between the glass plates or quartz plates to solidify. And used as a gel.

[0003]

In the conventional gel electrophoresis plate as described above, it takes time to prepare, and urea contained in the gel is deposited from the side end surface of the gel electrophoresis plate on which the laser enters. Irradiation light is scattered or refracted due to the drying of the gel, and there is a problem that laser irradiation may not be successful. Furthermore, it is inconvenient that only a certain distance from the migration start point can be irradiated, and it is not possible to select various migration distances, and the width direction of the gel migration plate (lateral direction)
There was also a problem that the temperature distribution of could not be made uniform and the practically usable range was extremely narrow. The present invention has been made in view of the above circumstances, and an object of the present invention is to solve the above-mentioned problems in the conventional technique and provide a highly practical gel electrophoresis tank for photodetection. .

[0004]

The above object of the present invention comprises at least two buffer liquid tanks and a gel electrophoresis plate composed of a gel sandwiched between glass plates and the like, and a liquid junction is provided between them. In the gel electrophoresis tank for light detection, the gel electrophoresis tank for light detection is characterized in that a side end surface portion of the gel electrophoresis plate is partially covered with the buffer solution tank.

[0005]

In the photodetection gel electrophoresis tank according to the present invention, the electrophoresis upper and lower buffer solution tanks and the electrophoresis plate holder are integrated, and the gel electrophoresis plate is housed in a box made of a transparent material. By creating a gel in the state, a wide area on the side surface can be used as an irradiation window, and the gel can be created by a simple operation. That is, a buffer solution tank-integrated electrophoretic plate holder made of a transparent material such as an acrylic plate is placed with the electrophoretic plate holding surface facing upward, and two glass plates are placed on the electrophoretic plate holder with a spacer interposed therebetween to migrate Tighten with a plate fixture and inject gel to solidify. With this configuration, it is not necessary to provide an irradiation window or to seal the surroundings and prevent the gel from flowing out, unlike the conventional case. The transparent side plate of this buffer solution tank functions as a window for laser light irradiation, and the lower buffer solution tank prevents excess gel material from leaking to the outside.

[0006]

Embodiments of the present invention will now be described in detail with reference to the drawings. FIG. 1 is a perspective view showing a state in which a buffer liquid tank-electrophoresis plate-integrated electrophoresis tank, which is an embodiment of the present invention, is longitudinally cut, and FIG. 2 is an exploded view thereof. In both figures, 1 is a buffer liquid tank forming plate, which has a partition plate 1a in the middle thereof,
The partition plate 1a divides the buffer solution tank into an upper buffer solution tank 1A and a lower buffer solution tank 1B. A step is formed on the side plate 1b of the buffer liquid tank forming plate 1 described above, and a gel electrophoresis plate is fitted therein as described later. Further, the side plate 1b of the buffer liquid tank forming plate 1 contains packings 8 and 9 for separating the upper buffer liquid tank 1A and the lower buffer liquid tank 1B by the action of the gel electrophoresis plate. U-shaped and inverted U-shaped packing rubber grooves are provided. 2 is a migration plate
(A) and 3 show electrophoretic plates (B). By forming gel between these electrophoretic plates, which will be described later, by pressing it against the packing groove surface of the buffer liquid tank forming plate 1 described above. , Two separated buffer solution tank-electrophoresis plate integrated electrophoresis tanks are formed. As shown in FIG. 2, in the buffer liquid tank-electrophoresis plate integrated electrophoresis tank shown in this embodiment, the buffer liquid tank forming plate 1, the electrophoretic plate (A) 2, the spacer 1
1, the migration plate (B) 3 and the holding plate 5 are stacked in this order, and the holding plate 5 and the buffer solution tank forming plate 1 are fastened with screws 7.
The above-mentioned holding plate 5 is provided with two packings 10 for contacting and sealing the surface of the buffer solution and the migration plate to prevent the buffer solution from leaking.

A method of using the buffer solution tank-electrophoresis plate integrated electrophoresis tank configured as described above will be described below. First, the electrophoresis tank assembled so that the electrophoresis plate (B) 3 is on the lower side is placed at an angle, and the gel material is injected into the lower buffer solution tank 1B. It enters and then solidifies to form a gel for electrophoretic separation. Excess gel material accumulates in the lower buffer solution tank 1B, but eventually solidifies. There is no need to spill around and require cleaning as in the conventional case. After the gel is solidified, the buffer solution is poured into the upper buffer solution tank 1A and the lower buffer solution tank 1B and set in the measuring device. Lower buffer solution tank 1
One or two holes for injecting the buffer solution are provided in the upper part of B, and the solution can be injected from here. The migration plate (A) 2 in contact with the buffer solution is shorter than the migration plate (B) 3, and the gel thus formed can contact the buffer solution. Electrophoresis plate
(B) 3 and the integrated buffer solution tank 1A form a container. Upper buffer liquid tank 1A and lower buffer liquid tank 1B
Electrophoresis is performed by applying a voltage to the linear electrode 6 installed therein. The electrophoretic current flows through the gap between both electrophoretic plates and the gel formed between the buffer solution tank and the side surface of the electrophoretic plate. The excitation laser passes through the side plate of the buffer solution tank and irradiates the gel through the gap between the spacers. Since the gel is formed in close contact with the acrylic side plate 1b of the buffer liquid tank forming plate 1 described above, there is no inconvenience such as lens action due to swelling of the gel at the incident portion. In addition, since the airtightness is good, urea which is present in a large amount in the gel is not deposited in the vicinity of the light incident part, and there is no inconvenience that the transmittance of irradiation light is lowered. The fluorescence image obtained is received by a line sensor or a two-dimensional detector, and DN
Separately detect A and the like.

Regarding the processing of the detected signal, etc., the method described in the above-mentioned Bio / Tech 6 , 816-821, 1988 etc. may be used. According to the above-described embodiment, the gel electrophoresis tank for photodetection is a buffer solution tank-electrophoresis plate integrated type, so that the labor of gel preparation is simple, and the light incident part can be stably and easily secured. In addition, it is easy to carry, and a wider effective utilization area than before can be taken on the electrophoresis plate.
In the past, this is due to heat dissipation from the end surface of the migration plate,
Even if the cm migration plate was used, the temperature was uniform only in the central area of about 10 cm, and it could not be effectively utilized, whereas in the buffer solution tank-migration plate integrated migration tank according to this example, both ends were The part is covered with an acrylic plate, and since the current flows through the gel formed on the side end, the temperature at the side end does not drop and the so-called smileing phenomenon does not occur as in the past. is there. Furthermore, since the buffer solution tank is in contact with the glass surface over a wide range, the temperature uniformity is better, and it can be sufficiently utilized for electrophoretic separation up to about 2 cm from the side end. For this reason, when a migration plate with a width of 20 cm is used, it can be utilized for migration separation over a central portion of 16 cm or more.

[0009]

As described above in detail, according to the present invention, in addition to the advantages that the preparation of the gel is easy, the light incident portion can be stably and easily secured, and in addition to being easy to carry, This has the remarkable effect of realizing a migration tank capable of providing a large effective utilization area on the migration plate.

[0010]

[Brief description of drawings]

FIG. 1 is a cross-sectional assembly view of a buffer solution tank-electrophoresis plate integrated electrophoresis tank.

FIG. 2 is an assembly view of a buffer solution tank-electrophoresis plate integrated electrophoresis tank.

[Explanation of symbols]

1: buffer tank, 2: migration plate A, 3: migration plate B, 4:
Gel, 5: Electrophoresis plate holding plate, 6: Electrophoresis voltage application electrode,
7: screw, 8: packing for separating the upper buffer liquid tank,
9: Packing for separating the lower buffer liquid tank, 10: Packing for preventing leakage, 11: Spacer.

Claims (3)

[Claims] 1. A photo-detection gel electrophoresis tank comprising at least two buffer liquid tanks and a gel electrophoresis plate composed of a gel sandwiched between glass plates or the like. A gel electrophoresis tank for photodetection, characterized in that a side end surface portion of the gel electrophoresis plate is covered with a part of the buffer solution tank. 2. The gel electrophoresis tank for photodetection according to claim 1, wherein at least a part of a portion of the buffer solution tank which is in contact with the side end surface portion of the gel electrophoresis plate is made of a transparent material. . 3. The gel electrophoresis tank for photodetection according to claim 1, wherein the buffer solution tank is separated by a glass plate or the like holding the gel to form two buffer solution tanks. .