JP3244786U - agarose gel filled bottle - Google Patents

Abstract

An object of the present invention is to provide an agarose gel-filled bottle for preparing an agarose gel for electrophoresis by a simple method while ensuring uniformity of agarose concentration. SOLUTION: An agarose gel-filled bottle 1 includes a container body 3 filled with agarose dissolved in an electrophoresis buffer in a gelled state 4, and a lid body 2, the capacity of the agarose gel being: It is 1/2 or less of the capacity of the container body. The agarose gel can be redissolved by heating in a microwave oven with the lid removed or loosened. [Selection diagram] Figure 1

Description

The present invention relates to an agarose gel-filled bottle in which agarose dissolved in an electrophoresis buffer is filled in a container made of plastic or the like so that the gelled agarose can be redissolved.

Gel electrophoresis is frequently used as a method for separating and analyzing proteins and DNA fragments because it is relatively simple and provides high-resolution results. Gel materials used for gel electrophoresis include agarose and polyacrylamide, but agarose gel is particularly frequently used because it is easier to prepare and non-toxic than polyacrylamide gel. has been done.

Agarose gel is generally attached to a horizontal slab type submarine electrophoresis device, and to prevent the surface of the gel from drying out during electrophoresis, it is immersed in a running buffer to the extent that the surface of the gel is covered. It is being done. Due to its molecular structure, agarose has a larger network structure when gelled compared to acrylamide, so it has a small molecular sieving effect and is said to be unsuitable for separating low molecules. Furthermore, when dissolving agarose in an electrophoresis buffer, if the heated solution is not sufficiently mixed, agarose powder or solid matter may remain, resulting in a non-uniform gel.

In order to detect clear bands in agarose gel electrophoresis, it is necessary to prepare an agarose gel with an appropriate concentration according to the molecular weight of the sample to be separated, and also to ensure that the agarose concentration is uniformly distributed throughout the gel or locally. This is very important.

Patent Document 1 discloses a microwave heating beverage filling bottle that can be safely heated in a microwave oven. The label has a structure in which the outer periphery of the bottle container body is covered with a label, and the label is provided with a strip-shaped window in the vertical direction through which the capacity of the beverage present in the bottle container body can be visually seen, and a strip-shaped window is provided along the window. At intervals, the output and heating time of the microwave oven are displayed in accordance with the volume of beverage present in the bottle main body. However, a redissolved agarose gel filled in a microwaveable bottle has not yet been commercialized.

Utility model registration No. 3208002

If the uniformity of the agarose gel for electrophoresis is not sufficient, there is a problem in that sufficient resolution cannot be obtained, especially for relatively small DNAs of about several hundred bases. Therefore, an object of the present invention is to provide an agarose gel-filled bottle for preparing an agarose gel for electrophoresis by a simple method while ensuring uniformity of agarose concentration.

The present invention was made to solve the above problems, and is based on the knowledge that the above problems can be solved by redissolving an agarose gel that has been previously dissolved and solidified in an electrophoresis buffer. That is, the agarose gel-filled bottle of the present disclosure includes a container body filled with gelled agarose dissolved in an electrophoresis buffer, and a lid, and the capacity of the agarose gel is equal to the capacity of the container body. It is characterized by being 1/2 or less.

According to the present invention, an experimenter can prepare an agarose gel for electrophoresis using a simple method.

FIG. 1 is a side view illustrating an example of an agarose gel-filled bottle according to the present disclosure. FIG. 1 is a perspective view illustrating an example of an agarose gel-filled bottle according to the present disclosure.

Hereinafter, an agarose gel-filled bottle according to the present disclosure will be described based on a specific embodiment. FIG. 1 is a side view of an agarose gel-filled bottle 1 according to the present disclosure. This agarose gel-filled bottle 1 has a lid 2 and a container body 3, and the container body 3 is filled with agarose gel 4. Agarose gel 4 is a gelled state of agarose dissolved in an electrophoresis buffer. As used herein, agarose is a neutral polysaccharide isolated and purified from red algae (Amanita). More specifically, it refers to a polysaccharide consisting of a repeating structure of alternating bonds of 1→3-linked β-D-galactose and 1→4-linked 3,6-anhydro-α-L-galactose. It is said that small amounts of sulfonic acid esters, sulfuric acid esters, pyruvic acid ketals, carboxyl groups, etc. are present at the 6- and 6-positions (Electrophoresis 1999, 20, 1455-1461). In the present invention, commercially available agarose can be suitably used. For example, Agarose S manufactured by Nippon Gene Co., Ltd. can be mentioned.

The electrophoresis buffer is not particularly limited, and may include Tris-acetic acid-ethylenediaminetetraacetic acid buffer (TAE buffer), Tris-boric acid-ethylenediaminetetraacetic acid buffer (TBE buffer), or Tris-glycine buffer (TG buffer solution). Here, "Tris" means trishydroxymethylaminomethane. Furthermore, the pH of the gel-preparing buffer is not particularly limited, but may be from 7.0 to 8.5. In addition, the above-mentioned TAE buffer, TBE buffer, or TG buffer contained in the electrophoresis buffer contains the same salt component (for example, Tris, etc.) for gel preparation and for the electrode solution used in the electrophoresis tank. The concentration of the salt component contained in the electrode solution may be lower than the concentration of the salt component contained in the gel preparation buffer.

Agarose has a large difference between its melting temperature and gelation temperature, and the gelation temperature range is 32 to 45°C, and the melting temperature is usually 80 to 95°C. Therefore, agarose can be easily gelled by melting it in a buffer at 80 to 95°C, injecting it into the container body 3, and leaving it at room temperature. The capacity of the container body 3 is not particularly limited, but is 100 mL to 1 L, preferably 200 mL to 500 mL, and more preferably about 250 mL, considering ease of heating in a microwave oven, autoclave, etc. The volume of the agarose gel 4 filled in this container body 3 is preferably 2/3 or less, more preferably 1/2 or less of the volume of the container body 3. By setting the capacity of the agarose gel 4 in this way, it is possible to prevent boiling over due to bumping when heated in a microwave oven. The lower limit of the capacity of the agarose gel 4 to be filled may be about 1/10 of the capacity of the container body 3, and is most preferably about 100 mL.

The material of the container body 3 is not particularly limited, and may include various types of glass bottles, as well as plastic materials such as PC (polycarbonate resin), PP (polypropylene resin), and PE (polyethylene resin). Plastic materials are suitable from the viewpoint of ease of molding. Moreover, it is preferable that the container body 3 is made of transparent plastic. However, the term "transparent" includes translucent and colored transparency, and it is sufficient that the content of the agarose gel can be seen through the container body 3 to the extent that the volume of the agarose gel can be confirmed.

The material of the lid body 2 is also not particularly limited, but from the viewpoint of sealing properties after agarose gel is injected, a plastic material with mechanical strength is preferable, and examples include PC (polycarbonate resin), PET (polyethylene terephthalate resin), and POM (polyacetal resin). ), PTFE (fluororesin), PEEK (polyetheretherketone), PF (phenol resin), EP (epoxy resin), etc. can be used.

FIG. 2 is a perspective view of the agarose gel-filled bottle 1 according to the present disclosure. The shape of the container body 3 may be any shape as long as it has a cylindrical shape with a bottom, the upper end of which is narrowed, and has a neck 6 and an opening 5. It is optional to provide ribs and the like as appropriate. The height of the container body 3 is preferably 120 mm to 250 mm. As long as it is 120 mm or more, the filling capacity of the agarose gel is not too small. On the other hand, if it is 250 mm or less, it can be easily heated in a household microwave oven. From this viewpoint, the height of the container body 3 is more preferably 130 mm to 200 mm, especially 150 mm or more or 190 mm or less, even more preferably 160 mm or more or 180 mm or less, and even more preferably 165 mm or more or 175 mm or less. It is particularly preferable that

The opening 5 of the container body 3 is preferably a wide-mouthed bottle from the viewpoint of suppressing the pressure increase inside the container due to heating and preventing the agarose solution from boiling over. Specifically, the diameter L1 of the opening 5 is at least 1/2 of the maximum diameter L2 of the body of the container body 3 (L1≧1/2*L2), and more preferably at least 2/3. (L1≧2/3*L2) is preferable. Thereby, the redissolved agarose gel can be easily cast and molded onto a gel production table. The agarose concentration used here can be appropriately selected depending on the purpose of use. Generally, a range of 0.8 to 3.0% by mass is preferred, and a range of about 2% by mass is particularly preferred. Since the agarose gel is uniformly dissolved, even molecular weight marker DNA of several hundred base pairs can be separated individually using about 2% by mass gel.

[Method for manufacturing agarose gel filled bottle]
(1) Preparation of TBE and TAE buffer The TBE buffer consists of 10.8 g of tris(hydroxymethyl)aminomethane [manufactured by Nacalai Tesque], 0.75 g of sodium ethylenediaminetetraacetic acid dihydrogen dihydrate [manufactured by Nacalai Tesque]. ] in 1 L of distilled water, and then 5.5 g of boric acid (manufactured by Nacalai Tesque). The TAE buffer was prepared by dissolving 4.84 g of tris(hydroxymethyl)aminomethane (manufactured by Nacalai Tesque) and 0.372 g of sodium ethylenediaminetetraacetic acid dihydrogen dihydrate (manufactured by Nacalai Tesque) in 1 L of distilled water. It was prepared by dissolving 0.412 g of sodium acetate (manufactured by Nacalai Tesque).

(2) Preparation of agarose gel 2 g of agarose (Agarose S, Nippon Gene Co., Ltd.) was weighed into a 250 mL product bottle, 100 mL of TAE buffer was added, and the mixture was dissolved in a microwave oven (700 W). It boiled in about a minute, so I took it out of the microwave and stirred it by hand. This process (putting in the microwave → boiling → taking out and stirring) was repeated three times, and it was visually confirmed that the agarose had completely melted. The agarose gel was left to stand at room temperature for 1 hour, and it was determined that the agarose gel had solidified, so it was transferred to a refrigerator at 4°C and solidified well.

Alternatively, a heating method using an autoclave may be used. Agarose and TAE buffer were weighed out in the same manner as above and dissolved in an autoclave (105°C, 1 minute). Thereafter, when the temperature of the autoclave reached 90° C., the autoclave was taken out and stirred by hand, and it was determined that the agarose had been dissolved. Thereafter, the agarose was solidified by "standing at room temperature for 1 hour → 4°C for 1 hour" in the same manner as above.

[effect]
(1) An agarose gel-filled bottle 1 comprising a container body 3 filled with gelled agarose dissolved in an electrophoresis buffer, and a lid 2, in which the capacity of the agarose gel is equal to the capacity of the container body. It was set to be 1/2 or less.
Therefore, the agarose gel-filled bottle of the present invention allows an experimenter to prepare an agarose gel for electrophoresis using a simple method.

(2) The agarose gel can be redissolved by removing or loosening the lid 2 and heating it in a microwave oven. Therefore, a gel in which agarose is uniformly dissolved can be produced.

(3) The diameter of the opening of the container body was set to be 1/2 or more of the maximum diameter of the body of the container body 3. Therefore, it is possible to prevent the re-dissolved agarose from boiling over, and to cast the re-dissolved agarose gel on a gel production stand and easily mold it.

(4) The container body 3 was made transparent to the extent that the capacity of the agarose gel could be confirmed. Therefore, the remaining amount of agarose gel can be easily confirmed.

(5) The concentration of the agarose gel was 0.8 to 3% by mass. Therefore, an agarose gel with an appropriate concentration can be produced according to the molecular weight of the sample.

1 Agarose gel filled bottle 2 Lid 3 Container body 4 Agarose gel 5 Opening 6 Neck

Claims (5)

An agarose gel-filled bottle comprising a container body filled with gelled agarose dissolved in an electrophoresis buffer, and a lid, wherein the capacity of the agarose gel is 1/2 of the capacity of the container body. Below is an agarose gel filled bottle. The agarose gel-filled bottle according to claim 1, wherein the agarose gel can be redissolved by heating in a microwave oven with the lid removed or loosened. The agarose gel-filled bottle according to claim 1 or 2, wherein the diameter of the opening of the container body is 1/2 or more of the maximum diameter of the body of the container body. The agarose gel-filled bottle according to claim 1 or 2, wherein the container body is transparent enough to confirm the capacity of the agarose gel. The agarose gel-filled bottle according to claim 1 or 2, wherein the agarose gel has a concentration of 0.8 to 3% by mass.

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