JPH0588296U - Device for recovering nucleic acid or protein from electrophoresis gel - Google Patents

Abstract

(57) [Summary] [Objective] The present invention is intended to recover a nucleic acid or a protein from an electrophoretic gel through a semipermeable membrane. In a device for recovering a nucleic acid or protein from a gel, a chamber containing an electrophoresis buffer, an electrode arranged in the chamber, a membrane blocking an opening of the chamber, or the inside of the chamber is provided. Nucleic acid from an electrophoretic gel, which is integrally formed of a membrane that divides into an electrode chamber and a collection chamber, the membrane being permeable to an electrophoretic buffer but impermeable to nucleic acids or proteins. Or a protein recovery device.

Description

[Detailed description of the device]

[0001]

[Industrial applications]

 The present invention relates to an apparatus for recovering a nucleic acid or a protein from an electrophoretic gel for the purpose of recovering a nucleic acid or a protein, particularly a nucleic acid from a band in an electrophoretic gel formed by a nucleic acid or a protein.

[0002]

[Prior 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, and the method for recovering the desired nucleic acid or protein from this band group is largely The following four types are 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 the electrophoresis is completed, 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. A cut is made in the vicinity of the migration direction of the desired band, DEAE cellulose paper is inserted into the cut, and electrophoresis is performed to adsorb the nucleic acid in the band to the membrane. Remove the paper, elute the nucleic acid adsorbed on the paper with a high concentration of salt, and perform ethanol precipitation on the eluate to recover the nucleic acid.

A method using low melting point agarose. A desired band is cut out from the gel, 1 to 5 times the amount of a buffer solution is added to the gel piece, the gel is melted at 65 ° C., treated with phenol, and then the aqueous layer containing the 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, buffer 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]

[Problems to be solved by devising]

 The known method has the following problems.

That is, is characterized by being able 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.

Cannot be applied to polyacrylamide gel, which is widely used along with agarose gel.

[0011] In the collected solution, gel pieces and the like are likely to be mixed, which may hinder the enzymatic reaction.

[0012]

[Means for Solving the Problems]

 The present invention relatively easily recovers nucleic acid or protein by applying a voltage through a membrane made of a material that is permeable to electrophoresis buffer but impermeable to nucleic acid or protein. Was successful.

That is, the present invention relates to a nucleic acid or protein recovery device from a gel, a chamber accommodating the buffer solution for electrophoresis, an electrode arranged in the chamber, a membrane for blocking an opening of the chamber, or the like. Is integrally formed with a membrane that divides the interior of the chamber into an electrode chamber and a collection chamber. The membrane is permeable to an electrophoretic buffer solution but impermeable to nucleic acids or proteins, such as cellulose. It is a device for recovering nucleic acid or protein from an electrophoretic gel, which is characterized by being made of a semipermeable membrane.

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 proteins, 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 electrode (counter electrode) paired with the electrode is electrically connected to the electrode through the gel. After applying the device, the gel and the counter electrode of the device, the electrophoresis buffer is recovered on the membrane or in the recovery chamber after the application is completed.

[0016]

First Embodiment Hereinafter, an embodiment of the present invention will be described 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, an electrophoretic buffer solution is contained in a hollow cylindrical body 5 that is integrally molded, and an electrode 3 is disposed in this 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 is opposed to 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 end 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. 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 100 V. After a few minutes, the application is stopped and the device of the present invention is separated 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]

Embodiment 2 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 therefore are omitted from the figure.

The structure of the device of the present invention in this embodiment is the same as that in FIG. 1 except that the recovery chamber 7 and the cap 8 constituting the recovery chamber 7 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 the same as in FIG.

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. Since the electrophoresis buffer containing the nucleic acid in the band 10 is attached to the surface of the semipermeable membrane 6 by the surface tension, the buffer is gently collected with a micropipette or the like, and the next step (for example, analysis) is performed. ) Etc.

[0024]

[Effect of the device]

 According to the device of the present invention, there is no need to perform operations (slicing, cutting, lysing, crushing, etc.) on the gel itself, and 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 or the like, it can be used for the next work without any concentration or purification operation after collection. There is an effect that can be transferred.

The device of the present invention saves the trouble of performing electrophoresis in a gel slice or a dialysis tube as compared with the method in the above conventional method, and there is no gel contamination, and the gel to the semipermeable membrane is removed. The time required for transfer of nucleic acid or protein is extremely short. Compared with the other methods, the operation is simple and the recovery rate is high. 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 the feature that there is no risk of gel fragments 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 electrode 4 electrode chamber 5 hollow cylinder 6 semipermeable membrane 7 collection chamber 9 gel 10 band

Claims (2)

[Scope of utility model registration request] 1. A device for recovering a nucleic acid or protein from a gel, a chamber containing an electrophoretic buffer, an electrode arranged in the chamber, a membrane for blocking an opening of the chamber, or the inside of the chamber. Is integrally formed with a membrane that divides the electrode chamber and a collection chamber into two parts, and the membrane is made of a material that is permeable to an electrophoresis buffer solution but not permeable to nucleic acids or proteins. A device for recovering nucleic acids or proteins from electrophoretic gels. 2. The apparatus for recovering nucleic acid or protein from an electrophoretic gel according to claim 1, wherein the material which is permeable to the buffer for electrophoresis but impermeable to the nucleic acid or protein is a semipermeable membrane.