This paper describes an improved immersion vertical plate electrophoresis tank with a cooling device. Instead of a commercially available compressible U-shaped rubber sleeve, a fixing frame and a spacer for controlling the thickness of the gel are made of plexiglass which does not deform. Secure the glass plate with screws. Sealed with a medical silicone tube to completely separate the positive and negative electrode running buffers. Tested for protein and nucleic acid electrophoresis, the effect is very good.
Electrophoresis is one of the most commonly used methods for biochemical and molecular biological experimental analysis. Electrophoresis is often required to be carried out at a constant temperature. Although there are various imported electrophoresis tanks with constant temperature cooling devices, they are expensive, each one is expensive. Thousands to more than 10,000 yuan. Although the domestic vertical plate electrophoresis tank also has a cooling device, the thickness of the prepared rubber sheet is generally only three, which cannot be adjusted at will, and often the inside of the sample hole is caused by the thickness of the comb being inconsistent with the thickness of the rubber sleeve spacer. There are residual gels that affect the loading and electrophoresis effects. Moreover, the thickness of the rubber septum is affected by the degree of tightness of the rotating screw when the two halves are fixed. To this end, in the work we refer to a variety of existing vertical electrophoresis tank style, designed an immersed vertical plate electrophoresis tank, simple, inexpensive and good results.
1 production of electrophoresis tank
The pattern of the electrophoresis tank is shown in the attached drawings. The whole groove is made of plexiglass, the outer shell is a rectangular groove, the size is 90×150×230cm (can be reduced as needed), and a fixing frame is arranged in the middle to fix the glass plate and separate the electrophoresis tank into positive and negative electrodes. . The specific structure of the fixing frame is that two pieces of plexiglass sheets each having a width of 20 mm, a length of 230 arms and a thickness of 5 mm or more are bonded to each other on the entire length of the central portions of the left and right side walls of the short groove, and the two pieces are separated by 20 mm. The front two pieces are drilled with three ribbed holes, and the bottom two pieces are connected by a 15mm high plexiglass strip to form a single piece, which is u-shaped and blends with the bottom of the groove to prevent liquid leakage.
2 installation and filling
When installing the glass plate, the electrophoresis tank is laid flat, the plexiglass sheet with holes on the holder is facing upward, and the u-shaped frame is under. A medical silicone tube is placed on top of the U-shaped frame, and is also made into a u-shaped shape. A glass plate of 210×150 mm size is placed thereon, and the bottom of the plate extends into the bottom of the electrophoresis tank, and two spacers are placed on both sides of the plate. Each 10-15 x 230 mm, the thickness depends on the thickness of the gel required for electrophoresis. Place a glass plate on the spacer, the size is 225 × 150mm, and the bottom of the plate is about 3mm from the bottom of the electrophoresis tank. Finally, put the plastic screw on the rib hole of the fixing frame and gradually tighten it. At this time, the silicone tube is gradually flattened and pressed, and the two glass plates are also fixed. The whole electrophoresis tank is divided into two grooves: front and rear. The groove is the positive electrode and the rear groove is the negative electrode. The glass tube is placed in each of the two slots, and the constant temperature water is passed to maintain the constant temperature of the running buffer and the gel immersed therein. When filling the polyacrylamide gel, the bottom of the glass plate is sealed with agarose gel, or sealed by other methods, and then glued as usual.
3 Discussion
The advantage of the immersion vertical plate electrophoresis tank is that the buffer tank is equipped with a constant temperature cooling water device, so that the electrophoresis buffer can be kept at a certain set temperature, and the electrophoresis gel plate is immersed in the buffer, and the gel is generated during electrophoresis. The heat is carried away by the buffer so that the electrophoresis can be carried out at a constant temperature. Therefore, the electrophoresis effect is better than that of an electrophoresis tank without a thermostat. The domestic vertical plate sandwich-cooled electrophoresis tank sold in the market has the above advantages, but the thickness of the gel plate is determined by the thickness of the rubber spacer, and the latter slightly changes with the size of the fixed force; more, the rubber spacer The thickness of the strip is often slightly different from the thickness of the toothed comb. Generally, the toothed comb is slightly thinner. Thus, there is often a small amount of residual gel in the well of the filled gel, so that the sample is not added. Neat, seriously affecting the electrophoresis effect. In view of this, we designed the above electrophoresis tank. When the polyacrylamide gel plate is filled, the gel thickness is controlled by a spacer made of plexiglass or other plastic, and the toothed comb and the spacer can be cut by the same plate. Therefore, the thickness is uniform, the sample hole is well formed, and the gel plates of different thicknesses can be prepared by randomly selecting the plates of different thicknesses as the spacers and the combs.
We use a tightening screw to fix and press the glass plate and the spacer. When the gel plate is filled, no liquid leaks. At the same time, a flexible silicone tube is placed on the back three sides of the lower glass plate. The screws are also pressed against the silicone tube to completely separate and seal the two buffer tanks, and no short circuit occurs during electrophoresis. The above device was tested for protein electrophoresis and single-strand conformation polymorphism (SSCP) analysis of DNA, and the effect was excellent.
(BIOLOGY Zheng Feng, Zeng Shan, etc.)
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