1,Preparation of 1% agarose gel(70ml for big gel,50ml for small gel):Weigh 0.7 g(0.5 g) agarose in a conical flask, add 70 ml(50ml) 1×TAE, invert the mouth of the flask into a small beaker.

Microwave heating and boiling 3 times until all the agarose melted, shake well, that is, 1.0% agarose gel solution.

2, gel plate preparation: take the electrophoresis tank in the plexiglass inner tank (gel-making tank) washed, dried, and put into the gel-making glass plate. Take the transparent tape to seal the glass plate with the edge of both ends of the inner tank to form a mold.

Place the inner groove in a horizontal position and put the comb in a fixed position, mix the agarose gel solution cooled to about 65 ℃ and carefully pour it into the inner groove glass plate, so that the gel slowly unfolds until the entire surface of the glass plate to form a uniform gel layer.

3, leave it at room temperature until the gel completely solidified, pull the comb vertically and gently, remove the tape, and put the gel and the inner tank into the electrophoresis tank. Add 1×TAE electrophoresis buffer until the gel plate is not covered.

Extended information:

Principle of electrophoresis

Electrophoresis is a process whereby electrophoretic paint is deposited on the surface of a workpiece by the action of the cathode and anode, under the action of a voltage applied, and the ions of the charged paint move to the cathode and form an insoluble substance with the action of alkaline substances produced on the surface of the cathode.

It includes four processes:

1. Electrolysis

(Decomposition) at the cathode reaction initially for the electrolytic reaction, generating hydrogen and hydroxide ions OH, this reaction causes the cathode surface to form a highly alkaline boundary layer, when the cation and hydroxide role of insoluble substances, the coating film is deposited, the equation is: H2O → OH + H.

2. Electrophoretic movement

(Swimming, migration) cation resin and H + under the action of the electric field, to the cathode, while the anion to the anode moving process.

3. Electrodeposition

(Precipitation) On the surface of the coated workpiece, the cationic resin and the cathode surface alkaline action, neutralization and precipitation of non-deposited material, deposited on the coated workpiece.

4. Electro-osmosis

(Dehydration) The coating film on the surface of paint solid and workpiece is translucent with most capillaries, water is exuded from the cathode coating film, and under the action of electric field, it causes the coating film to be dehydrated, and the coating film is adsorbed on the surface of the workpiece, and completes the whole electrophoresis process.

Basic principle

Biological macromolecules such as proteins, nucleic acids, polysaccharides, etc. mostly have cationic and anionic groups, called amphoteric ions. Often dispersed in solution as particles, their electrostatic charge depends on the H+ concentration of the medium or interaction with other macromolecules in the electric field, the charged particles migrate to the cathode or anode, the direction of migration depends on the sign of their charge, this migration phenomenon is known as electrophoresis.

If a colloidal solution of a biomolecule is placed in an undisturbed electric field, the driving force that gives the particles a constant rate of migration is the effective charge on the particles, Q, and the potential gradient, E. They are in opposition to the frictional resistance of the medium, f. In a free solution this resistance is in opposition to the frictional resistance of the medium, F. In a free solution this resistance obeys Stokes' law.

Here v is the speed of movement of a particle of radius r in a medium with viscosity η. However, in gels, this resistance does not exactly obey Stokes' law. f depends on other factors in the medium, such as gel thickness, particle size, and even the endosmosis of the medium.

Electrophoretic mobility (mbility) m is specified as the distance d over which the particles migrate in time t under the influence of a potential gradient E.

Differences in mobility provide the basis for separating substances from mixtures, and the distance migrated is proportional to the mobility.

Baidu Encyclopedia - Electrophoresis