JPH081496Y2 - Device for recovering nucleic acid or protein from electrophoresis gel - Google Patents

Description

[Detailed description of the device]

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a device for recovering nucleic acid or protein from an electrophoretic gel, particularly for recovering nucleic acid or protein from a band in an electrophoretic gel formed by nucleic acid or protein. .

[0002]

2. Description of the Related Art When a biological component containing a nucleic acid or a protein is subjected to electrophoresis, the nucleic acid or the protein is separated due to the difference in the molecular weight to form a band. The desired nucleic acid or the protein is recovered from this band group. The following four types of methods are roughly known.

A method of migrating to a dialysis tube by electrophoresis. The desired band is cut out from the gel, the gel piece is placed in a dialysis tube together with a buffer solution, and the dialysis tube is energized between the electrodes of the electrophoresis apparatus to release the nucleic acid or protein in the buffer solution. After completion of electrophoresis, the buffer solution in the dialysis tube is taken and concentrated, and the nucleic acid is recovered by phenol extraction and ethanol precipitation.

A method of migrating to DEAE cellulose paper by electrophoresis. Make a cut near the migration direction of the desired band, insert DEAE cellulose paper into the cut,
Electrophoresis is performed to adsorb the nucleic acid in the band to the membrane. The paper is taken out, the nucleic acid adsorbed on the paper is eluted with a high concentration of salt, and the eluate is subjected to ethanol precipitation to recover the nucleic acid.

A method using low melting point agarose. A desired band is cut out from the gel, 1 to 5 volumes of a buffer solution is added to the gel piece, the gel is melted at 65 ° C., phenol treatment is performed, and then an aqueous layer containing a nucleic acid is recovered.

A method of crushing a gel. The desired band is cut out from the gel, the gel piece is mechanically disrupted, a buffer solution is added and left for several hours, then the gel fragment is removed by centrifugation to obtain a supernatant, and the nucleic acid is recovered from this supernatant. To do.

[0007]

The above-mentioned known method has the following problems.

That is, is characterized in that it is possible to obtain a wide range of molecular weights, but it is rarely used because it is an extremely complicated method, and a large molecule (nucleic acid having a length of about 5 kb or more is used. ) Is an effective method only for taking out.

The method (1) requires operations such as elution of nucleic acid from DEAE cellulose paper with a high-concentration salt solution and ethanol precipitation, and the recovery rate is not so good.

Not applicable to polyacrylamide gel, which is widely used along with agarose gel.

In the case of the above, gel fragments and the like are liable to be mixed in the collected solution, and the enzyme reaction may be hindered.

[0012]

The present invention is relatively easy to apply a voltage through a membrane made of a material that is permeable to an electrophoresis buffer solution but impermeable to nucleic acids or proteins. It succeeded in recovering nucleic acid or protein.

That is, the present invention provides a nucleic acid isolated in a gel.
Recover protein bands by gel electrophoresis
In the device, inside the chamber containing the electrophoresis buffer solution +
Electrodes are installed, and the electrophoresis buffer passes through the tip of the chamber.
However, even semipermeable membranes that do not allow nucleic acids or proteins to permeate
The volume and opening of the electrode chamber are extremely small and removable.
It is divided into a collection chamber and a grip is provided at one end of the chamber.
Integrated recovery body, and-gel with exposed electrode on the surface
Rukoto a and a stand is a recovery device of the nucleic acid or protein from the electrophoretic gel characterized. Another device
Shows the separated nucleic acid or protein bands in the gel.
Electrophoresis in a device that collects by gel electrophoresis
The + electrode is placed inside the chamber containing the buffer solution for
Although the buffer solution for electrophoresis permeates through the portion, nucleic acid or protein is passed through.
Cover the opening of the electrode chamber with a semipermeable membrane that does not allow white matter to pass through.
A collection body with a grip provided at one end of the chamber
And-consisting of a gel platform with the electrode exposed on the surface
Of nucleic acids or proteins from electrophoresis gels characterized by
It is a recovery device.

Examples of the buffer for electrophoresis include buffers usually used in electrophoresis, such as Tris-acetate buffer, Tris-phosphate buffer and Tris-borate buffer when recovering nucleic acids. When recovering the protein, Tris-glycine buffer or the like is used.

When recovering nucleic acid or protein from a gel using the device of the present invention, the present invention is designed so that the electrode (counter electrode) forming a pair with the electrode is electrically connected to the electrode through the gel. After arranging the apparatus, the gel and the counter electrode, the application is performed, and after the application is completed, the electrophoresis buffer on the membrane or in the recovery chamber is recovered.

[0016]

Embodiment 1 An embodiment of the present invention will be described below with reference to the drawings.

FIG. 1 shows an example of an apparatus for recovering nucleic acid using the apparatus of the present invention.

In FIG. 1, a hollow cylindrical body 5 integrally formed.
An electrophoretic buffer solution is housed therein, and the electrode 3 is arranged therein to form an electrode chamber 4. The opening of the electrode chamber 4 is partitioned by a semipermeable membrane 6 made of cellulose, and the lower end of the electrode chamber 4 faces a recovery chamber 7 filled with the same electrophoresis buffer solution as described above. The recovery chamber 7 is formed by detachably fitting a cap 8 to the non-end portion of the hollow cylindrical body 5.

The hollow cylindrical body 5 is inserted and attached to the grip portion 1 formed in a shape that is easy to hold by hand (to form a recovery body).
) . The gel 9 is placed on a plate 11 made of an ultraviolet-transparent acrylic resin on which an electrode 12 is arranged so that the band 10 to be collected is directly above the electrode 12. The electrode 3 and the electrode 12 are connected to the anode and the cathode of the power supply 2, respectively.

When recovering the nucleic acid from the gel 9, the device of the present invention is installed so that the opening of the recovery chamber 7 abuts on the band 10, and then the power supply 2 applies a voltage of 100V. After a few minutes, the application is stopped and the device according to the invention is released from the band 10. The electrophoresis buffer containing the nucleic acid in the band 10 is recovered from the recovery chamber 7 with a micropipette or the like and used in the next step (for example, analysis).

[0021]

Second Embodiment Another embodiment of the device of the present invention is shown in FIG.
In this figure, the gripping part and the power source are the same as those in FIG. 1, and are therefore omitted from the figure.

The structure of the device of the present invention in this embodiment is the same as that of FIG. 1 except that the recovery chamber 7 and the cap 8 constituting it in FIG. 1 are not provided. The arrangement of the plate 11, the electrode 12, the gel 9 and the like other than the device of the present invention is shown in FIG.
Is the same as

When recovering the nucleic acid from the gel 9, the device of the present invention is installed so that the semipermeable membrane 6 contacts the band 10, and then the power supply 2 applies 100 V. After a few minutes, the application is stopped and the device according to the invention is released from the band 10. The electrophoresis buffer containing the nucleic acid in band 10 is
Since it adheres to the surface of the semipermeable membrane 6 by surface tension, this buffer solution is gently collected with a micropipette or the like and used in the next step (for example, analysis).

[0024]

The device of the present invention is easy to operate and
It is easy, and there is no need to perform operations (slicing, cutting, lysing, crushing, etc.) on the gel itself , and the nucleic acid or protein can be recovered from the gel simply and in a short time. Moreover, since the recovered nucleic acid or protein solution has a very high concentration and is highly pure with no gel fragments and the like, the next work is performed without performing concentration and purification operations after recovery. There is an effect that can be.

The device of the present invention saves the labor of performing electrophoresis by putting it in a gel slice or a dialysis tube as compared with the method in the conventional method, and there is no gel contamination.
It takes a very short time to transfer the nucleic acid or protein from the gel to the semipermeable membrane. Compared with the other method,
Easy operation and high recovery rate. Compared to the following method, nucleic acid or protein can be recovered from any gel without being limited to agarose gel. Compared with the above method, it has a feature that there is no risk of gel pieces and the like being mixed in.

[Brief description of drawings]

FIG. 1 is a front view in which a part of an implementation device of the present invention is shown in section.

FIG. 2 is a front view in which a part of another embodied device is also shown in section and a part thereof is omitted. .

[Explanation of symbols]

 3, 12 Electrodes 4 Electrode chamber 5 Hollow cylinder 6 Semipermeable membrane 7 Collection chamber 9 Gel 10 Band

Continuation of the front page (51) Int.Cl. ⁶ Identification number Office reference number FI technical display location C12M 1/00 A G01N 33/50 P

Claims (2)

[Scope of utility model registration request] 1. A nucleic acid or protein separated in a gel.
In a device that collects the bands of gel by gel electrophoresis
Then , the + electrode is arranged inside the chamber containing the electrophoresis buffer,
The electrophoresis buffer passes through the tip of the chamber, but the nucleic acid
A semipermeable membrane that does not allow proteins to permeate
A removable collection chamber with a very small volume and opening.
It is a single piece that is cut and integrated with a grip at one end of the chamber.
And Osamu body, - electrode recovery device nucleic acids or proteins from electrophoresis gels wherein Rukoto such from the gel rests exposed on the surface of. 2. Nucleic acid or protein separated in gel
In a device that collects the bands of gel by gel electrophoresis
Then , the + electrode is arranged inside the chamber containing the electrophoresis buffer,
The electrophoresis buffer passes through the tip of the chamber, but the nucleic acid
Open the electrode chamber with a semipermeable membrane that does not allow proteins to permeate.
The mouth is covered and a grip is provided at one end of the chamber for integration.
From the recovery body and the gel mounting table with the electrode exposed on the surface
A device for recovering a nucleic acid or a protein from an electrophoretic gel, which comprises: