JP3360270B2 - Transfer device for elution of nucleic acid in gel - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION In general physics and chemistry, especially in molecular biology research, nucleic acids having different molecular weights and base sequences are separated by agarose gel or polyacrylamide gel electrophoresis and then eluted from agarose gel or polyacrylamide gel. Experiments have been widely conducted in which the data is transferred to a filter paper or a special film such as nitrocellulose or nylon for analysis. The present invention relates to a novel device for performing this efficiently.

[0002]

2. Description of the Related Art In a conventional elution transfer method, besides a method utilizing a capillary phenomenon using blotting paper or the like as shown in FIG. 3, an electrophoresis method, a forced passage of a buffer solution (salt solution) by decompression or pressurization. There is a method to make it.

[0003]

However, in the method utilizing the capillary phenomenon, a large amount of filter paper or blotting paper is required to allow a required amount of liquid to pass therethrough, and they are cut into appropriate sizes. It takes a lot of trouble to do it.
Further, there is a problem that the elution requires a long time as much as 6 to 20 hours, and the weight placed on the gel to promote close contact between the members deforms the gel, so that accurate transfer cannot be expected.

The electrophoresis method, the pressurization method and the depressurization method can perform elution transfer in a shorter time (within one hour) than the electrophoresis method. A high pressure pump and an expensive special sealing device are required for the pressurizing method and the depressurizing method, and there is a problem that each of them requires a considerable cost. Further, in the decompression method, there is a problem that the gel is apt to be deformed, and it is difficult to expect accurate elution transfer.

[0005]

According to the present invention, there is provided a pad tank in which a pad material of an appropriate size can be detachably mounted at a central portion, and a bottomless gap is provided at the central portion on the pad tank. Two horizontal buffer tanks are vertically movable, and both ends are immersed in a buffer solution filled in the two buffer tanks. Filter paper or a special membrane such as nitrocellulose or nylon and a gel such as agarose gel or polyacrylamide gel containing nucleic acid on the filter paper or special membrane are laminated on the pad material. The hanging water supply material is brought into contact with the upper surface of the gel, and the buffer solution is passed by utilizing the capillary action and gravity of the pad material to elute the nucleic acids in the gel, and is transferred to the filter paper or special membrane. So as to, it is intended to solve such problems.

[0006]

The present invention utilizes both capillary action and gravity,
A device that elutes and transfers nucleic acids from a gel in a short time.That is, by making the flow of a buffer solution for nucleic acid elution down, gravity is added to the capillary phenomenon, and the elution speed can be remarkably accelerated. At the same time, the weight is not required to prevent the gel from being deformed. The pad material is a material that can be restored by washing with water after use, and can be used repeatedly and repeatedly.

[0007]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS An embodiment will be described below with reference to the drawings.

FIG. 1 shows an apparatus according to an embodiment of the present invention. The bottom plate 1a is made of a material such as acrylic resin, and a rectangular frame composed of four side walls 1b is mounted and mounted on the bottom plate 1a. Tank 1 for opening the container, a rectangular frame 2 having the same depth as the pad tank 1, and bottom plates 3 on both sides of the rectangular frame 2 on the longitudinal side.
a, 3a and the inner tanks 3b, 3b, which are inclined upwardly, are formed into two layers of buffer tanks 3, 3. 4
Is a bottomless gap at the center located between the buffer tanks 3 and 3. The vertical cylinders 5 are mounted on the bottom plate 1a at the four outer corners of the pad tank 1 and can be detached from the vertical cylinders 5 so that the columns 6 can be freely detached.
Is fixed by screws 7 with knobs, and mounting cylinders 8 provided at the relative positions of the square frame 2 are fitted to the respective posts 6 so as to be movable up and down.
It can be stopped and fixed with.

Numeral 10 is a sponge-like pad material which is porous and excellent in water absorption, has a square or rectangular shape larger than a gel described later, and is formed with a smooth upper surface. Place it on the top so that it can be removed. The pad material 10 preferably has a certain degree of hardness and can be reused by washing and drying.

Reference numeral 11 denotes both ends immersed in a buffer solution to be filled in the buffer tanks 3 and 3, and a middle portion having no bottom gap 4.
This is a water supply material made of a band-shaped filter paper made of a material having a high capillary action efficiency formed with a length that can be hung down toward the pad material 10.

The gel 12 used for electrophoresis of nucleic acids is made of a material such as agarose gel or polyacrylamide gel.
It is approximately square or rectangular and formed in an appropriate size.

The gel 12 after the transfer of the nucleic acid by electrophoresis is placed on the pad material 10 with a filter paper for nucleic acid transfer or a special film 13 such as nitrocellulose or nylon interposed therebetween. The middle part of the water supply material 11 whose both ends are immersed in the liquid is suspended from the bottomless gap 4 to form a square frame 2
The middle part of the water supply material 11 is uniformly brought into contact with the upper surface of the gel 12 by the vertical movement adjustment.

Then, the buffer solution penetrates the water supply material 11 by capillary action, elutes the nucleic acid from the water supply material 11 while passing through the gel 12, and transfers the nucleic acid to the transfer filter paper or the special membrane 13 immediately below. However, the flow of the buffer solution reaching the pad material 10 acts very quickly because of the strong and almost uniform capillary phenomenon drawn by the pad material 10 having excellent water absorption, and furthermore, the gravitational force cooperates with the flow. Is completed in a short time within one hour.

After the operation is completed, the filter paper or the special membrane 13 to which the nucleic acid has been transferred is carried to the next experimental site, and the pad tank 1
The inside and the removed pad material 10 are washed with water, washed and dried,
The buffer solution tanks 3 and 3 are filled with a new buffer solution to prepare for the next operation.

The device can be easily disassembled by removing the square frame 2 by loosening the screw 9 with the knob and removing the support 6 by loosening the screw 7 with the knob.
The cleaning work can be made very easy, and when not in use, it can be stored compactly.

[0016]

As described above, according to the present invention, in an apparatus for eluting and transferring nucleic acids in a gel, accurate and rapid elution transfer of nucleic acids can be performed without causing deformation of the gel. Is generated. Moreover, this type of experiment can be performed more economically without the need for expensive special equipment and power consumption, and using as little consumables as blotter paper and filter paper. Produces an effect.

[Brief description of the drawings]

FIG. 1 is a perspective view showing an embodiment of the device of the present invention.

FIG. 2 is a sectional view showing an embodiment of the method of the present invention using the same device.

FIG. 3 is a diagram showing a conventional example of a method using a capillary phenomenon.

[Explanation of symbols]

 1a is a bottom plate 1b is a side wall plate 1 is a pad tank 2 is a square frame 3a, 3a is a bottom plate 3b, 3b is an inner wall 3, 3 is a buffer tank 4, a bottomless gap 5 is a vertical cylinder 6, a column 7 is a knob 7 Screws 8 are mounting cylinders 9 Screws with knobs 10 Pad materials 11 Water supply materials made of band-shaped filter paper 12 Gels 13 Filter paper for transfer or special membrane

Continuation of the front page (56) References BioTechniques, August 24, 1993, Vol. 15, No. 2, 260, 262 Analytical Biochemistry, February 14, 1992, Vol. 201, No. 1,134-139 (58) Field surveyed (Int. Cl. ⁷ , DB name) G01N 27/447 JICST file (JOIS)

Claims (1)

(57) [Claims] 1. A pad tank in which a pad material of an appropriate size can be detachably mounted at a central portion, and two horizontal buffer tanks having a bottomless gap at a central portion on the pad tank. An intermediate portion of a water supply material such as filter paper having a high capillary action efficiency in which both ends are immersed in a buffer solution filled in the two buffer tanks so as to be vertically movable is provided so as to hang down from the bottomless gap. A filter paper or a special membrane such as nitrocellulose or nylon and a gel such as an agarose gel or a polyacrylamide gel containing nucleic acid are laminated on the filter paper or the special membrane on the pad material. The nucleic acid in the gel is eluted by contacting the supplied water supply material and passing the buffer solution by utilizing the capillary action and gravity of the pad material, and the buffer solution is transferred to the filter paper or the special membrane. A nucleic acid elution transfer device in gel.