JP2835426B2 - Electrophoresis device - Google Patents
Electrophoresis deviceInfo
- Publication number
- JP2835426B2 JP2835426B2 JP7197498A JP19749895A JP2835426B2 JP 2835426 B2 JP2835426 B2 JP 2835426B2 JP 7197498 A JP7197498 A JP 7197498A JP 19749895 A JP19749895 A JP 19749895A JP 2835426 B2 JP2835426 B2 JP 2835426B2
- Authority
- JP
- Japan
- Prior art keywords
- electrophoresis
- buffer
- tank
- plates
- plate
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 238000001962 electrophoresis Methods 0.000 title claims description 216
- 239000000872 buffer Substances 0.000 claims description 58
- 239000007853 buffer solution Substances 0.000 claims description 27
- 239000000499 gel Substances 0.000 claims description 22
- 238000001816 cooling Methods 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 7
- 238000005192 partition Methods 0.000 description 8
- 239000000758 substrate Substances 0.000 description 7
- 125000006850 spacer group Chemical group 0.000 description 6
- 238000000926 separation method Methods 0.000 description 5
- 108091005461 Nucleic proteins Proteins 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 1
- 238000010420 art technique Methods 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 239000000498 cooling water Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 1
- 229960005542 ethidium bromide Drugs 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 230000020169 heat generation Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- ZIUHHBKFKCYYJD-UHFFFAOYSA-N n,n'-methylenebisacrylamide Chemical compound C=CC(=O)NCNC(=O)C=C ZIUHHBKFKCYYJD-UHFFFAOYSA-N 0.000 description 1
- HFWWEMPLBCKNNM-UHFFFAOYSA-N n-[bis(hydroxyamino)methyl]hydroxylamine Chemical compound ONC(NO)NO HFWWEMPLBCKNNM-UHFFFAOYSA-N 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 239000003507 refrigerant Substances 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 229920003002 synthetic resin Polymers 0.000 description 1
- 239000000057 synthetic resin Substances 0.000 description 1
- UEUXEKPTXMALOB-UHFFFAOYSA-J tetrasodium;2-[2-[bis(carboxylatomethyl)amino]ethyl-(carboxylatomethyl)amino]acetate Chemical compound [Na+].[Na+].[Na+].[Na+].[O-]C(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CC([O-])=O UEUXEKPTXMALOB-UHFFFAOYSA-J 0.000 description 1
Landscapes
- Electrostatic Separation (AREA)
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、電気泳動装置に関
する。さらに詳しくは、核酸あるいは蛋白質を分離する
に際し、一回の泳動で、多数の検体を分離することがで
きる電気泳動装置に関する。[0001] The present invention relates to an electrophoresis apparatus. More specifically, the present invention relates to an electrophoresis apparatus capable of separating a large number of samples by one electrophoresis when separating nucleic acids or proteins.
【０００２】[0002]
【従来の技術】核酸あるいは蛋白質を電気泳動で分離す
る手法は、生命科学の分野で広く用いられてきた。特
に、近年の遺伝子工学の進歩、発展とともに、ゲノムの
構造、配列決定に際して、一度に多数の検体を解析した
い場合がある。2. Description of the Related Art Techniques for separating nucleic acids or proteins by electrophoresis have been widely used in the field of life science. In particular, with the progress and development of genetic engineering in recent years, it is sometimes desired to analyze a large number of specimens at once when determining the structure and sequence of a genome.
【０００３】従来、用いられてきた電気泳動装置は、上
部の陰極側の緩衝液槽と下部の陽極側の緩衝液槽との２
槽を有する、いわゆる２槽式の電気泳動装置である。従
来の電気泳動装置は、例えば、図３に示すように、上部
電気泳動緩衝液槽１１ａと下部電気泳動緩衝液槽１１ｂ
とから構成されている。上部電気泳動緩衝液槽１１ａ
は、低面１９ａと、低面１９ａに対して垂直な３つの側
壁１８ａ1〜１８ａ3と電気泳動板１３とで構成される。
電気泳動板１３は、側壁１８ａ1および１８ａ2とに設け
られた取付部１７ａに押圧手段１７により、側壁１８ａ
と1および１８ａ2とに密着して取り付けられる。このよ
うにして上部電気泳動緩衝液槽１１ａは、緩衝液を貯留
できるようになる。上部電気泳動緩衝液槽１１ａの低面
１９ａには電極１２ａが配置されている。上部電気泳動
緩衝液槽１１ａの内部には、側壁１８ａ1および１８ａ2
を貫通する緩衝液の冷却用コイル１５が配置されてお
り、電気泳動緩衝液を冷却する。下部電気泳動緩衝液槽
１１ｂの低面１９ｂには電極１２ｂが配置されている。
上部電気泳動緩衝液槽１１ａは、電気泳動緩衝液の界面
１４ａが、電気泳動板１３中の電気泳動用ゲル１３’の
上部１３’ａよりも上部にくるように、電気泳動緩衝液
で満たされる。他方、下部電気泳動緩衝液槽１１ｂは、
電気泳動板１３中の電気泳動用ゲル１３’の下部１３’
ｂよりも、電気泳動緩衝液の界面１４ｂが上部にくるよ
うに、電気泳動緩衝液で満たされている。このようにし
て、上部電気泳動緩衝液槽１１ａと下部電気泳動緩衝液
槽１１ｂとの間で電圧を印加して、電気泳動を行う。[0003] Conventionally used electrophoresis apparatuses are composed of a buffer solution tank on the upper cathode side and a buffer solution tank on the lower anode side.
This is a so-called two-tank electrophoresis apparatus having a tank. For example, as shown in FIG. 3, the conventional electrophoresis apparatus includes an upper electrophoresis buffer tank 11a and a lower electrophoresis buffer tank 11b.
It is composed of Upper electrophoresis buffer tank 11a
It has a low surface 19a, composed of the three side walls 18a 1 ~18a 3 electrophoretic plate 13 perpendicular to the lower surface 19a.
Electrophoresis plate 13, the pressing means 17 to a mounting portion 17a provided on the side wall 18a 1 and 18a 2, side walls 18a
When mounted in close contact with the first and 18a 2. In this way, the upper electrophoresis buffer solution tank 11a can store the buffer solution. The electrode 12a is arranged on the lower surface 19a of the upper electrophoresis buffer solution tank 11a. Inside the upper electrophoresis buffer tank 11a, side walls 18a 1 and 18a 2
A cooling coil 15 for a buffer solution that penetrates the electrophoresis buffer solution is arranged. The electrode 12b is arranged on the lower surface 19b of the lower electrophoresis buffer tank 11b.
The upper electrophoresis buffer tank 11a is filled with the electrophoresis buffer so that the interface 14a of the electrophoresis buffer is higher than the upper portion 13'a of the electrophoresis gel 13 'in the electrophoresis plate 13. . On the other hand, the lower electrophoresis buffer tank 11b is
Lower part 13 'of electrophoresis gel 13' in electrophoresis plate 13
It is filled with the electrophoresis buffer so that the interface 14b of the electrophoresis buffer is higher than b. In this way, electrophoresis is performed by applying a voltage between the upper electrophoresis buffer solution tank 11a and the lower electrophoresis buffer solution tank 11b.
【０００４】しかし、この電気泳動装置は、使用する電
気泳動ゲルの強度不足から大面積を有する電気泳動ゲル
を作成できないため、一度に多数の検体を処理すること
ができない。また、電気泳動板１３の対向する側壁１８
ａ3を、電気泳動板１３に変えて、２枚を同時に電気泳
動することも可能と考えられる。さらには、この装置を
横に並べて泳動板を多数配置し、多数の検体を同時に処
理することも可能と考えられる。しかし、多数の検体を
一度に処理するには、装置が大きくなりすぎて取扱いが
不便になり、装置自体も高価なものとなるという欠点が
ある。[0004] However, this electrophoresis apparatus cannot produce a large-area electrophoresis gel due to insufficient strength of the electrophoresis gel to be used, and therefore cannot process a large number of samples at once. Further, the opposing side walls 18 of the electrophoresis plate 13
The a 3, instead of the electrophoresis plate 13, is considered possible to simultaneously electrophoresed two. Furthermore, it is considered that this apparatus can be arranged side by side and a large number of electrophoresis plates are arranged to simultaneously process a large number of samples. However, in order to process a large number of specimens at once, there is a drawback that the apparatus becomes too large, handling becomes inconvenient, and the apparatus itself becomes expensive.
【０００５】従って、一度に多数の検体が電気泳動可能
な装置の開発が望まれていた。Therefore, there has been a demand for the development of an apparatus capable of electrophoresing a large number of specimens at one time.
【０００６】[0006]
【発明が解決しようとする課題】本発明は、上記従来の
電気泳動装置が有する上記欠点を解決することを目的と
するものである。SUMMARY OF THE INVENTION An object of the present invention is to solve the above-mentioned drawbacks of the conventional electrophoresis apparatus.
【０００７】[0007]
【課題を解決するための手段】本発明は、一槽型の電気
泳動装置であって、泳動緩衝液が収容されており、２以
上の泳動板を浸漬し得る泳動緩衝液槽；該泳動緩衝液槽
の底面に対して、該２以上の泳動板をほぼ垂直方向にも
しくは水平方向に平行に配置する手段；および、該泳動
緩衝液槽の対向する側壁あるいはその近傍にそれぞれ配
置される泳動電極であって、該一方の泳動電極は泳動緩
衝液槽の一側壁のほぼ上端部であって該泳動緩衝液の界
面下に配置され、他方の泳動電極は該一側壁と対向する
側壁の下端部に配置される、泳動電極；を有する、電気
泳動装置に関する。SUMMARY OF THE INVENTION The present invention relates to a one-tank type electrophoresis apparatus, in which an electrophoresis buffer is accommodated and in which two or more electrophoresis plates can be immersed; Means for arranging the two or more electrophoresis plates substantially vertically or horizontally in parallel with the bottom surface of the liquid tank; and electrophoresis electrodes respectively arranged on opposing side walls of the electrophoresis buffer tank or in the vicinity thereof Wherein the one electrophoresis electrode is disposed substantially at the upper end of one side wall of the electrophoresis buffer tank and below the interface of the electrophoresis buffer, and the other electrophoresis electrode is the lower end of the side wall facing the one side wall. , An electrophoresis device having a migration electrode.
【０００８】好適な実施態様においては、本発明の電気
泳動装置は、該２以上の泳動板の間に泳動用緩衝液が通
過可能な空間を有する。[0008] In a preferred embodiment, the electrophoresis apparatus of the present invention has a space between the two or more electrophoresis plates through which a buffer for electrophoresis can pass.
【０００９】好適な実施態様においては、本発明の電気
泳動装置の、泳動緩衝液槽の底面に対して、ほぼ垂直に
重ね合わせる泳動板あるいはほぼ水平に積層する泳動板
が、それぞれ、独立した泳動板である。In a preferred embodiment, in the electrophoresis apparatus of the present invention, an electrophoresis plate which is superimposed almost vertically on the bottom surface of the electrophoresis buffer tank or an electrophoresis plate which is superimposed almost horizontally on the bottom surface of the electrophoresis buffer tank are independent electrophoresis plates. It is a board.
【００１０】好適な実施態様においては、本発明の電気
泳動装置は、泳動緩衝液の循環手段を有する。[0010] In a preferred embodiment, the electrophoresis apparatus of the present invention has means for circulating an electrophoresis buffer.
【００１１】さらに、好適な実施態様においては、本発
明の電気泳動装置は、泳動緩衝液の冷却手段を有する。Further, in a preferred embodiment, the electrophoresis apparatus of the present invention has means for cooling an electrophoresis buffer.
【００１２】さらに本発明は、上記本発明の電気泳動装
置を用いて、一度に多数の検体を電気泳動する方法に関
する。Further, the present invention relates to a method for electrophoresing a large number of specimens at one time using the above-described electrophoresis apparatus of the present invention.
【００１３】泳動緩衝液が収容された泳動緩衝液槽内
に、泳動板を槽の底面に対してほぼ垂直状態で平行に配
置し、あるいはほぼ水平状態で平行に配置した後、電極
にて、電圧を印加すると、該各電極は該泳動緩衝液槽の
対向する側壁あるいはその近傍の上下部にそれぞれ配置
されていることにより、複数の泳動板には均一に電圧が
印加される。従って、多数のゲルを用いても、ほぼ均一
の電気泳動が可能となり、一度に多量の検体の電気泳動
が可能となる。[0013] In the electrophoresis buffer tank containing the electrophoresis buffer, the electrophoresis plate is arranged in a substantially vertical state in parallel with the bottom surface of the tank, or in an almost horizontal state in parallel, and then the electrode is When a voltage is applied, the electrodes are arranged on the opposing side walls of the electrophoresis buffer tank or on the upper and lower portions in the vicinity thereof, so that the voltages are uniformly applied to the plurality of electrophoresis plates. Therefore, even if a large number of gels are used, substantially uniform electrophoresis can be performed, and a large amount of specimen can be electrophoresed at one time.
【００１４】また、近接する各泳動板の間に泳動用緩衝
液が通過可能な空間を設けることにより、泳動用緩衝液
が各泳動板の側面を移動することが可能になる。従っ
て、この泳動用緩衝液を冷却水中を通過させて冷却する
ことにより、泳動板の過剰な発熱を防止して泳動の分離
能を向上させ得る。Further, by providing a space through which the electrophoresis buffer solution can pass between adjacent electrophoresis plates, the electrophoresis buffer solution can move on the side surface of each electrophoresis plate. Therefore, by passing the electrophoresis buffer through cooling water and cooling it, excessive heat generation of the electrophoresis plate can be prevented, and the electrophoretic separation ability can be improved.
【００１５】[0015]
【発明の実施の形態】本発明の電気泳動装置を図１に示
す。本発明の電気泳動装置は、一槽型の泳動緩衝液槽１
と、該泳動緩衝液槽１の底部に配置された電気泳動板３
を保持するための保持板８と、該泳動緩衝液槽１の側壁
部内面に配置されており泳動緩衝液槽１の底面９に対し
て、２以上の泳動板３をほぼ垂直方向に平行に配置する
押圧手段７と、該泳動緩衝液槽１内に満たされる緩衝液
の液面近くと、泳動緩衝液槽１の底部内にそれぞれ配置
される電極２ａ、２ｂと、泳動用緩衝液入口５および泳
動用緩衝液出口６と、を有している。FIG. 1 shows an electrophoresis apparatus according to the present invention. The electrophoresis apparatus of the present invention comprises a single-type electrophoresis buffer tank 1.
And an electrophoresis plate 3 arranged at the bottom of the electrophoresis buffer tank 1.
And a holding plate 8 for holding the electrophoresis buffer solution tank 1. The two or more electrophoresis plates 3 are arranged on the inner surface of the side wall portion of the electrophoresis buffer solution tank 1 and substantially parallel to the bottom surface 9 of the electrophoresis buffer solution tank 1 in a substantially vertical direction. Pressing means 7 to be arranged; electrodes 2a and 2b arranged near the liquid surface of the buffer solution filled in the electrophoresis buffer solution tank 1 and in the bottom of the electrophoresis buffer solution tank 1; And an electrophoresis buffer outlet 6.
【００１６】上記保持板８は、電気泳動槽１の底面９の
両側部にそれぞれ配置され、電気泳動槽１の底面９と間
隙を有する状態で電気泳動板３を下方から支持し得るも
のであれば、形状を問わない。あるいは、上記保持板８
は、電気泳動槽１の側壁部内面に配置され得る。上記保
持板８は電気泳動板３と嵌合し得る溝あるいは仕切を有
し得る。The holding plates 8 are respectively arranged on both sides of the bottom surface 9 of the electrophoresis tank 1 and can support the electrophoresis plate 3 from below with a gap with the bottom surface 9 of the electrophoresis tank 1. Any shape can be used. Alternatively, the holding plate 8
Can be arranged on the inner surface of the side wall of the electrophoresis tank 1. The holding plate 8 may have a groove or a partition that can be fitted with the electrophoresis plate 3.
【００１７】上記押圧手段７は、２以上の泳動板３を泳
動槽本体１の底面９に対して垂直方向に平行に保持し得
るものであればよく、複数使用し得る。泳動板３を垂直
方向に保持する押圧手段７としては、バネ、ゴム、合成
樹脂等の弾性部材やネジ等の取付部材が使用され得る。The pressing means 7 only needs to be able to hold two or more electrophoresis plates 3 in a direction perpendicular to the bottom surface 9 of the electrophoresis tank main body 1, and a plurality of them can be used. As the pressing means 7 for holding the electrophoresis plate 3 in the vertical direction, an elastic member such as a spring, rubber, or synthetic resin, or a mounting member such as a screw may be used.
【００１８】本実施例においては、２以上の泳動板をほ
ぼ垂直方向にもしくは水平方向に平行に配置する手段と
して、上記保持板８と押圧手段７を使用しているが、こ
れらには限定されないことはいうまでもない。例えば、
電気泳動槽１の側壁部内に、電気泳動板３の厚みの間隔
で、低面９に対して垂直に電気泳動板３と嵌合し得る仕
切あるいは溝を有し得る。この仕切あるいは溝があれ
ば、上記保持板８が電気泳動板３と嵌合し得る仕切ある
いは溝を有しなくてもよい。保持板８が電気泳動板３と
嵌合し得る溝あるいは仕切を有する場合には、特に、安
定に配置され得る。In this embodiment, the holding plate 8 and the pressing means 7 are used as means for arranging two or more electrophoresis plates substantially in a vertical direction or in a horizontal direction, but the present invention is not limited to these. Needless to say. For example,
In the side wall portion of the electrophoresis tank 1, there may be provided a partition or a groove which can be fitted to the electrophoresis plate 3 perpendicularly to the lower surface 9 at intervals of the thickness of the electrophoresis plate 3. If the partition or the groove is provided, the holding plate 8 does not need to have a partition or a groove that can be fitted to the electrophoresis plate 3. When the holding plate 8 has a groove or a partition that can be fitted with the electrophoresis plate 3, the holding plate 8 can be arranged particularly stably.
【００１９】また、電気泳動板３は、電気泳動槽１の外
部から、電気泳動槽１の底面９に対して垂直方向に立設
するように、カセット式に嵌合して組立られ得、カセッ
ト毎、電気泳動層１に浸漬され得る。この場合でも、上
記カセットは、カセットの低面に対して垂直に電気泳動
板３を配置し得る手段、例えば前記押圧手段７、あるい
はカセットの側面部内に溝または仕切を有し得る。The electrophoresis plate 3 can be assembled in a cassette manner from the outside of the electrophoresis tank 1 so as to stand vertically from the bottom surface 9 of the electrophoresis tank 1. Each time, it can be immersed in the electrophoresis layer 1. Even in this case, the cassette may have means for arranging the electrophoresis plate 3 perpendicular to the lower surface of the cassette, for example, the pressing means 7 or a groove or a partition in the side surface of the cassette.
【００２０】上記泳動電極２は、それぞれ電気泳動槽１
内の対角する位置に配置されている。すなわち、該一方
の泳動電極２は、該泳動緩衝液槽１の側壁あるいはその
近傍において泳動緩衝液４の界面下に配置され、他方の
泳動電極２は、該側壁に対向する側壁において電気泳動
槽１の底部近傍位置に配置されている。この場合、各電
極２は電気泳動板３のゲルの厚み方向の側壁にそれぞれ
配置されている。このように泳動電極２をそれぞれ対向
する側壁の上下端部に配置することにより、電圧が各泳
動板（ゲル）３に対して、均一に印加されるので、各ゲ
ル間での試料の分離のばらつきが少ない。Each of the electrophoresis electrodes 2 is connected to an electrophoresis tank 1
Are arranged at diagonally opposite positions. That is, the one electrophoresis electrode 2 is disposed below the interface of the electrophoresis buffer solution 4 at or near the side wall of the electrophoresis buffer solution tank 1, and the other electrophoresis electrode 2 is disposed on the side wall opposing the side wall. 1 is located near the bottom. In this case, each electrode 2 is arranged on the side wall of the gel of the electrophoresis plate 3 in the thickness direction. By arranging the electrophoresis electrodes 2 at the upper and lower ends of the opposing side walls in this manner, a voltage is uniformly applied to each electrophoresis plate (gel) 3, so that the sample separation between the gels can be performed. There is little variation.
【００２１】上記泳動用緩衝液入口５および泳動用緩衝
液出口６は、緩衝液の冷却装置（図示せず）と連結され
ており、緩衝液４を冷却し得る。緩衝液４を冷却するこ
とにより、電気泳動中の発熱が吸収され、電気泳動の精
度が向上する。泳動用緩衝液入口５は、好ましくは、電
気泳動槽１のいずれかの側壁の下方に、泳動用緩衝液出
口６は、冷却等の効率等を考慮すると、電気泳動槽１の
該側壁と対向する側の側壁の上方に配置され得る。The electrophoresis buffer inlet 5 and the electrophoresis buffer outlet 6 are connected to a buffer cooling device (not shown) so that the buffer 4 can be cooled. By cooling the buffer solution 4, the heat generated during the electrophoresis is absorbed, and the accuracy of the electrophoresis is improved. The electrophoresis buffer inlet 5 is preferably located below one of the side walls of the electrophoresis tank 1, and the electrophoresis buffer outlet 6 is opposed to the side wall of the electrophoresis tank 1 in consideration of efficiency such as cooling. May be located above the side wall of the side.
【００２２】なお、泳動用の緩衝液４は、緩衝液４自体
を循環して冷却する場合の他、冷却用冷媒を循環させる
冷却パイプ等を電気泳動槽１内に通過させることによ
り、冷却され得る。The electrophoresis buffer solution 4 is cooled by circulating the buffer solution 4 itself and cooling it, or by passing the electrophoresis tank 1 through a cooling pipe or the like for circulating a cooling refrigerant. obtain.
【００２３】上記電気泳動板３は、それぞれ独立して用
いられ得、あるいは一体としても用いられ得るが、作業
の効率、電気泳動の分離能等を考慮すると、各泳動板３
は独立している方が好ましく、電気泳動板３の間を緩衝
液が移動できる構造であることがより好ましい。最も好
ましい電気泳動板３を図２で説明する。The above-mentioned electrophoresis plates 3 can be used independently or can be used as a single unit. However, in consideration of work efficiency, electrophoresis separation ability, etc., each of the electrophoresis plates 3 can be used.
Are preferably independent, and more preferably have a structure in which a buffer solution can move between the electrophoresis plates 3. The most preferred electrophoresis plate 3 will be described with reference to FIG.
【００２４】図２（ｂ）は、基板２０、２０の両端部間
にスペーサー２１、２１を配置し、両基板２０、２０間
で形成される空間内にゲル２５を充填して構成される電
気泳動板３の底面図である。この電気泳動板３の一方の
基板２０の両端部にはさらに別のスペーサー２１が取り
付けられている。FIG. 2B shows an electric structure in which spacers 21 and 21 are arranged between both ends of the substrates 20 and 20 and a gel 25 is filled in a space formed between the substrates 20 and 20. FIG. 4 is a bottom view of the electrophoresis plate 3. Further spacers 21 are attached to both ends of one substrate 20 of the electrophoresis plate 3.
【００２５】図２（ａ）の上側の図は、図２（ｂ）の上
側に示されている、基板２０の両端部にスペーサー２１
が取り付けられている電気泳動板３の正面図であり、コ
ーム２９は試料を負荷するための溝を形成するために使
用する。図２（ａ）の下側の図は、電気泳動板３の片方
の基板２１を取り除いた状態の底面図である。電気泳動
板３は基板２０の両端にスペーサー２１を有している。
このスペーサー２１は、ゲルが保持される必要な長さを
有していればよい。The upper part of FIG. 2A shows spacers 21 on both ends of the substrate 20 shown in the upper part of FIG.
FIG. 2 is a front view of the electrophoresis plate 3 to which a sample is attached, and a comb 29 is used to form a groove for loading a sample. 2A is a bottom view of the electrophoresis plate 3 from which one substrate 21 has been removed. The electrophoresis plate 3 has spacers 21 at both ends of a substrate 20.
The spacer 21 only needs to have a length necessary to hold the gel.
【００２６】図２（ｃ）は、上記図２（ｂ）で示した電
気泳動板３を一つのユニットとして、複数重ねた状態の
底面図である。各ユニットを重ねることにより隣接する
電気泳動板３間に空間２７が生じている。このようにし
て生じた空間２７は、緩衝液の移動が自由であり、電気
泳動中の発熱が緩衝液により吸収されて、電気泳動の精
度も向上する。FIG. 2C is a bottom view showing a state in which the electrophoresis plate 3 shown in FIG. A space 27 is created between adjacent electrophoresis plates 3 by stacking the units. In the space 27 thus generated, the buffer solution can move freely, and the heat generated during the electrophoresis is absorbed by the buffer solution, so that the accuracy of the electrophoresis is also improved.
【００２７】本発明の装置は、泳動緩衝液槽１の底面９
に対して、２以上の泳動板３をほぼ水平に積層して配置
され得る。この場合、上記泳動緩衝液槽１の底面９に対
して垂直に電気泳動板３を配置する場合と同様に、泳動
電極２の一方は、泳動緩衝液槽１のある壁面の上端部で
あって泳動緩衝液の界面４下に、泳動電極２の他方は、
該壁面と対向する壁面の下端部に配置される。The apparatus according to the present invention comprises a bottom surface 9 of the electrophoresis buffer tank 1.
On the other hand, two or more electrophoresis plates 3 can be arranged so as to be substantially horizontally stacked. In this case, as in the case where the electrophoresis plate 3 is arranged perpendicular to the bottom surface 9 of the electrophoresis buffer tank 1, one of the electrophoresis electrodes 2 is located at the upper end of the wall surface where the electrophoresis buffer tank 1 is located. Under the interface 4 of the electrophoresis buffer, the other of the electrophoresis electrodes 2
It is arranged at the lower end of the wall facing the wall.
【００２８】電気泳動板３（ゲル）を水平に配置する手
段としては、上記した保持板８、押圧手段７、あるい
は、電気泳動槽１の側壁部内に、電気泳動板３の厚みの
間隔で、低面９に対して水平に電気泳動板３と嵌合し得
る仕切あるいは溝を有し得る。また、電気泳動板３は、
電気泳動槽１の外部から、電気泳動槽１の底面９に対し
て水平方向に配置し得るように、カセット式に嵌合して
組立られ得、カセット毎、電気泳動層１に浸漬され得
る。この場合でも、上記カセットは、カセットの低面に
対して水平に電気泳動板３を配置し得る手段、例えば前
記押圧手段７、あるいはカセットの側面部内に溝または
仕切を有し得る。電気泳動板３としては、上記と同じ電
気泳動板３が用いられ得、好適には、図２の構造を有す
る電気泳動板３が用いられ得る。As means for horizontally arranging the electrophoresis plate 3 (gel), the above-mentioned holding plate 8, the pressing means 7, or the inside of the side wall of the electrophoresis tank 1, at intervals of the thickness of the electrophoresis plate 3, are used. It may have a partition or a groove that can be fitted to the electrophoresis plate 3 horizontally with respect to the lower surface 9. In addition, the electrophoresis plate 3
The cassette can be fitted and assembled from the outside of the electrophoresis tank 1 so as to be arranged in a horizontal direction with respect to the bottom surface 9 of the electrophoresis tank 1, and each cassette can be immersed in the electrophoresis layer 1. Also in this case, the cassette may have a means for arranging the electrophoresis plate 3 horizontally with respect to the lower surface of the cassette, for example, the pressing means 7 or a groove or a partition in the side surface of the cassette. As the electrophoresis plate 3, the same electrophoresis plate 3 as described above can be used, and preferably, the electrophoresis plate 3 having the structure of FIG. 2 can be used.
【００２９】さらに、水平に配置する場合、試料の負荷
のための溝は、当業者に周知のコームが使用され得、試
料が負荷できるように電気泳動板をずらして配置し得
る。[0029] Further, in the case of the horizontal arrangement, the groove for loading the sample may be a comb well known to those skilled in the art, and the electrophoresis plate may be staggered so that the sample can be loaded.
【００３０】[0030]
【実施例】縦、横それぞれ10ｃｍ、厚さ１ｍｍの分離ゲ
ルを１２枚作成した。ゲルは、4-10％のポリアクリルア
ミドゲル（アクリルアミド：ビスアクリルアミド＝19:
1）を緩衝液（４５ｍＭトリスヒドロキシアミノメタ
ン、４５ｍＭほう酸、１ｍＭエチレンジアミン４酢酸ナ
トリウム、ｐＨ８．３）中で作成した。分離ゲルを有す
る電気泳動板を１ｍｍの間隔をおいて、電気泳動槽の低
面にほぼ垂直に、電気泳動槽内に配置した。電気泳動槽
内を上記と同じ組成の泳動用緩衝液で満たし、ΦX174フ
ァージのＤＮＡを制限酵素HaeIIIで消化したものを、12
枚のゲルにチャージし、２２℃、200Vの定電圧で泳動し
た。泳動後、１mlあたり0.5μgのエチジウムブロマイド
を含む電気泳動緩衝液を用いてゲルを染色し、320nmの
波長の紫外線を照射してＤＮＡの分離を確認した。上下
の泳動電極を結んだ線とゲルとの切片の角度が４５度以
下の場合、すべてのゲルで再現性のある均一な移動度を
示した。EXAMPLE Twelve separation gels each having a length and width of 10 cm and a thickness of 1 mm were prepared. The gel was a 4-10% polyacrylamide gel (acrylamide: bisacrylamide = 19:
1) was made in buffer (45 mM trishydroxyaminomethane, 45 mM boric acid, 1 mM sodium ethylenediaminetetraacetate, pH 8.3). An electrophoresis plate having a separation gel was placed in the electrophoresis tank at an interval of 1 mm and almost perpendicular to the lower surface of the electrophoresis tank. The inside of the electrophoresis tank was filled with an electrophoresis buffer having the same composition as above, and the DNA of the ΦX174 phage digested with the restriction enzyme
The gel was charged and electrophoresed at 22 ° C. and a constant voltage of 200V. After the electrophoresis, the gel was stained with an electrophoresis buffer containing 0.5 μg of ethidium bromide per 1 ml, and the DNA was confirmed by irradiation with ultraviolet light having a wavelength of 320 nm. When the angle between the line connecting the upper and lower electrophoresis electrodes and the section of the gel was 45 degrees or less, all gels showed uniform mobility with reproducibility.
【００３１】[0031]
【発明の効果】本発明の装置によれば、複数の泳動板に
実質的に均一な電圧を印加できるので、一度に多量の検
体の電気泳動が可能となる。According to the apparatus of the present invention, since a substantially uniform voltage can be applied to a plurality of electrophoresis plates, a large amount of specimens can be electrophoresed at one time.
【図１】本発明の一実施例である電気泳動装置１を示す
図である。FIG. 1 is a diagram showing an electrophoresis apparatus 1 according to one embodiment of the present invention.
【図２】本発明の電気泳動装置１に用いる好適な電気泳
動板を示す図である。FIG. 2 is a view showing a preferred electrophoresis plate used for the electrophoresis apparatus 1 of the present invention.
【図３】従来用いられている電気泳動装置を示す図であ
る。FIG. 3 is a view showing a conventionally used electrophoresis apparatus.
１ 一槽型泳動緩衝液槽 ２ 電極 ３ 電気泳動板 ４ 緩衝液 ５ 泳動用緩衝液入口 ６ 泳動用緩衝液出口 ７ 押圧手段 ８ 保持板 ９ 泳動緩衝液槽の底面 １１ 電気泳動緩衝液槽 １２ 電極 １３ 電気泳動板 １４ 電気泳動緩衝液の界面 １５ 冷却用コイル １７ 押圧手段 １８ 側壁 １９ 低面 ２０ 基板 ２１ スペーサー ２５ ゲル ２７ 空間 ２９ コーム DESCRIPTION OF SYMBOLS 1 Single electrophoresis buffer tank 2 Electrode 3 Electrophoresis board 4 Buffer 5 Electrophoresis buffer inlet 6 Electrophoresis buffer outlet 7 Pressing means 8 Holding plate 9 Electrophoresis buffer tank bottom 11 Electrophoresis buffer tank 12 Electrodes Reference Signs List 13 electrophoresis plate 14 interface of electrophoresis buffer solution 15 cooling coil 17 pressing means 18 side wall 19 low surface 20 substrate 21 spacer 25 gel 27 space 29 comb
フロントページの続き (56)参考文献 特開 昭55−110945（ＪＰ，Ａ) 特開 昭54−12897（ＪＰ，Ａ) 特開 平５−215714（ＪＰ，Ａ) 特開 平２−269935（ＪＰ，Ａ) 特開 平２−218941（ＪＰ，Ａ) 特開 昭62−184343（ＪＰ，Ａ) 特開 平４−143651（ＪＰ，Ａ) 特開 昭56−97860（ＪＰ，Ａ) 実開 昭49−103884（ＪＰ，Ｕ) 実開 昭55−25（ＪＰ，Ｕ) 実開 昭57−201961（ＪＰ，Ｕ) (58)調査した分野(Int.Cl.6，ＤＢ名) G01N 27/447 ＪＩＣＳＴファイル（ＪＯＩＳ)Continuation of front page (56) References JP-A-55-110945 (JP, A) JP-A-54-12897 (JP, A) JP-A-5-215714 (JP, A) JP-A-2-269935 (JP) , A) JP-A-2-21841 (JP, A) JP-A-62-184343 (JP, A) JP-A-4-143651 (JP, A) JP-A-56-97860 (JP, A) 49-103884 (JP, U) Fully open 1979-55 (JP, U) Fully open 1979-201961 (JP, U) (58) Fields investigated (Int. Cl. 6 , DB name) G01N 27/447 JICST file (JOIS)
Claims (6)
得る泳動緩衝液槽；該泳動緩衝液槽の底面に対して、該
２以上の泳動板をほぼ垂直方向にもしくは水平方向に平
行に配置する手段；および、 該泳動緩衝液槽の対向する側壁あるいはその近傍にそれ
ぞれ配置される泳動電極であって、該一方の泳動電極は
泳動緩衝液槽の一側壁のほぼ上端部であって該泳動緩衝
液の界面下に配置され、他方の泳動電極は該一側壁と対
向する側壁の下端部に配置される、泳動電極；を有す
る、電気泳動装置。1. An electrophoresis apparatus of a one-tank type, comprising an electrophoresis buffer tank containing an electrophoresis buffer and capable of immersing two or more electrophoresis plates; Means for arranging the two or more electrophoresis plates substantially in a vertical direction or parallel to a horizontal direction; and electrophoresis electrodes respectively arranged on or near opposing side walls of the electrophoresis buffer solution tank, wherein An electrophoresis electrode, wherein the electrode is disposed at substantially the upper end of one side wall of the electrophoresis buffer tank and below the interface of the electrophoresis buffer, and the other electrophoresis electrode is disposed at the lower end of the side wall opposite to the one side wall; An electrophoresis apparatus comprising:
が通過可能な空間を有する、請求項１に記載の電気泳動
装置。2. The electrophoresis apparatus according to claim 1, wherein a space through which an electrophoresis buffer can pass is provided between the two or more electrophoresis plates.
た泳動板から構成される、請求項１または２に記載の電
気泳動装置。3. The electrophoresis apparatus according to claim 1, wherein each of the two or more electrophoresis plates comprises an independent electrophoresis plate.
る、請求項１ないし３のいずれかの項に記載の電気泳動
装置。4. The electrophoresis apparatus according to claim 1, further comprising means for circulating an electrophoresis buffer.
る、請求項４に記載の電気泳動装置。5. The electrophoresis apparatus according to claim 4, further comprising means for cooling an electrophoresis buffer.
いて、一度に多数の検体を電気泳動する方法であって、
該方法は、 １槽型の電気泳動装置であって、 泳動緩衝液が収容されており、２以上の泳動板を浸漬し
得る泳動緩衝液槽；該泳動緩衝液槽の底面に対して、該
２以上の泳動板をほぼ垂直方向にもしくは水平方向に平
行に配置する手段；および、 該泳動緩衝液槽の対向する側壁あるいはその近傍にそれ
ぞれ配置される泳動電極であって、該一方の泳動電極は
泳動緩衝液槽の一側壁のほぼ上端部であって該泳動緩衝
液の界面下に配置され、他方の泳動電極は該一側壁と対
向する側壁の下端部に配置される、泳動電極；を有する
電気泳動装置に、２以上の泳動板を配置する工程、 検体をゲルに負荷する工程、を包含する、方法。6. A method for electrophoresing a large number of samples at once using an electrophoresis plate filled with two or more gels,
The method is a one-tank type electrophoresis apparatus, in which an electrophoresis buffer is accommodated and an electrophoresis buffer tank in which two or more electrophoresis plates can be immersed; Means for arranging two or more electrophoresis plates in a substantially vertical direction or in a horizontal direction; and electrophoresis electrodes arranged on or near opposing side walls of the electrophoresis buffer tank, wherein the one electrophoresis electrode Is a substantially upper end of one side wall of the electrophoresis buffer tank and is disposed below the interface of the electrophoresis buffer, and the other electrophoresis electrode is disposed at a lower end of the side wall facing the one side wall. A method comprising: arranging two or more electrophoresis plates on an electrophoresis apparatus, and loading a sample on a gel.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP7197498A JP2835426B2 (en) | 1995-08-02 | 1995-08-02 | Electrophoresis device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP7197498A JP2835426B2 (en) | 1995-08-02 | 1995-08-02 | Electrophoresis device |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0943196A JPH0943196A (en) | 1997-02-14 |
| JP2835426B2 true JP2835426B2 (en) | 1998-12-14 |
Family
ID=16375478
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP7197498A Expired - Lifetime JP2835426B2 (en) | 1995-08-02 | 1995-08-02 | Electrophoresis device |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2835426B2 (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3573130B2 (en) * | 1999-09-24 | 2004-10-06 | 独立行政法人農業生物資源研究所 | Highly efficient genome scanning |
| JP4572022B2 (en) * | 2000-07-06 | 2010-10-27 | 光夫 板倉 | Sample introduction apparatus for slab gel electrophoresis apparatus and sample introduction method thereof |
| JP5445234B2 (en) * | 2010-03-09 | 2014-03-19 | 凸版印刷株式会社 | Electrophoresis cassette and method for producing electrophoresis gel cassette |
| JP5375681B2 (en) * | 2010-03-09 | 2013-12-25 | 凸版印刷株式会社 | Electrophoresis cassette |
| US9234874B2 (en) * | 2011-02-24 | 2016-01-12 | Bio-Rad Laboratories, Inc. | Dimensional stabilization of slab gel cassettes to prevent distortion caused by swelling gels |
-
1995
- 1995-08-02 JP JP7197498A patent/JP2835426B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0943196A (en) | 1997-02-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US5865974A (en) | Apparatus and method for electrophoresis | |
| US6379516B1 (en) | Apparatus and method for electrophoresis | |
| US20070074973A1 (en) | Electrophoresis Apparatus For Simultaneous Loading of Multiple Samples | |
| EP0313293B1 (en) | Method and apparatus for use in biological testing | |
| US4622124A (en) | Device for horizontal electroblotting | |
| WO1998057161A1 (en) | Device for rehydration and electrophoresis of gel strips and method of using the same | |
| US4589965A (en) | Method for electroblotting | |
| US4391688A (en) | Electrophoresis system for multiple agarose slab gels | |
| JP2004510168A (en) | Electrophoresis system | |
| US20110011741A1 (en) | Apparatus and method for electrophoresis | |
| JPH0772117A (en) | High-resolution two-dimensional electrophoresis method and device for performing method thereof | |
| Svoboda et al. | Rapid electrotransfer of proteins from polyacrylamide gel to nitrocellulose membrane using surface-conductive glass as anode | |
| JPH09288091A (en) | Gene analyzer | |
| JP2835426B2 (en) | Electrophoresis device | |
| JP4213216B2 (en) | Methods and materials for optimizing electronic hybridization reactions | |
| US20100032296A1 (en) | Systems and methods for quantitative analyte transfer | |
| US6165337A (en) | Semi-dry electrophoresis apparatus and method | |
| Jones et al. | Electrophoresis tank for running multiple slab gels | |
| JP3541971B2 (en) | Analysis method using electrophoresis device | |
| CA1181034A (en) | Electrophoresis system for multiple agarose slab gels | |
| JPH0134114Y2 (en) | ||
| JPS58140634A (en) | Simple two-dimensional electrophoresis method | |
| Dunn | Electroblotting of proteins from 2-D polyacrylamide gels | |
| Heller et al. | Membrane electrophoresis of DNA | |
| JP2000028578A (en) | Plate-like electrode for electric transfer, and electric transfer method therefor |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 19980831 |
|
| S533 | Written request for registration of change of name |
Free format text: JAPANESE INTERMEDIATE CODE: R313533 |
|
| S533 | Written request for registration of change of name |
Free format text: JAPANESE INTERMEDIATE CODE: R313533 |
|
| R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
| EXPY | Cancellation because of completion of term |