How to use electrophoresis apparatus and electrophoresis tank
Electrophoresis instrument: Electrophoresis technology is an indispensable and important analytical method for molecular biology research. Electrophoresis is generally divided into two categories: free interface electrophoresis and zone electrophoresis. Free interface electrophoresis does not require supports, such as isoelectric focusing electrophoresis, constant velocity electrophoresis, density gradient electrophoresis, and microelectrophoresis. . For zone electrophoresis, various types of substances are used as supports. Commonly used supports include filter paper, cellulose acetate film, non-gel support, gel support, and silica gel-G thin layer. Molecular biology The most commonly used in the field is agarose gel electrophoresis. The so-called electrophoresis refers to the movement of charged particles in the electric field. Different substances have different speeds in the electric field due to the different charges and molecular weights. According to this feature, the electrophoresis method can be used to qualitatively or quantitatively analyze different substances. , Or a certain mixture for component analysis or single component extraction preparation, which has extremely important significance in clinical testing or experimental research. The electrophoresis instrument is designed and manufactured based on the above principles. The following is a brief introduction to its use and precautions.
Instructions
1. First connect the two electrodes of the electrophoresis tank to the DC output terminal of the electrophoresis instrument with a wire, and pay attention not to reverse the polarity.
2. The power switch of the electrophoresis instrument is turned to the off position, the voltage knob is turned to the minimum, and the voltage stabilizing current mode and the voltage and current range are selected according to the work needs.
3. Turn on the power, slowly rotate the voltage adjustment knob until the desired voltage is reached, and set the electrophoresis termination time, at which time electrophoresis begins.
4. After the work is completed, the knobs and switches should be turned to zero or off, and the electrophoresis plug should be pulled out.
Precautions
1. After the electrophoresis instrument is energized and enters the working state, it is forbidden for the human body to contact the electrodes, electrophoresis materials and Other parts that may be charged, nor to take things into the electrophoresis tank. If necessary, power off first to avoid electric shock. At the same time, the instrument must have a good ground terminal to prevent leakage.
2. After the instrument is powered on, do not temporarily add or remove the output lead plug to prevent short circuit. Although a fuse is attached inside the instrument, the short circuit may still cause damage to the instrument.
3. Because the resistance value of different media supports is different, the amount of current passed during electrophoresis is also different, and the swimming speed and the time required to reach the end are also different. Therefore, electrophoresis of different media supports should not be performed on the same electrophoresis apparatus at the same time.
4. When the total current does not exceed the rated current of the instrument (maximum current range), it can be used in association with multiple slots, but be careful not to overload, otherwise it will easily affect the life of the instrument.
5. In some special cases, when the electrophoresis input of the instrument needs to be checked, it is allowed to start with no load in a steady state, but it must be connected to the load before starting in a steady state, otherwise the pointer of the voltmeter will jump greatly, which is easy to cause unnecessary Of the man-made machine is damaged.
6. If abnormal phenomena such as loud noise, discharge or abnormal smell are found during use, the power supply must be cut off immediately and repaired to avoid accidents.
How to use the sandwich type vertical electrophoresis tank:
1. Components
(1) A pair of liquid storage tanks equipped with platinum wires, equipped with cooling devices and electrophoresis buffer outlets.
(B) The concave rubber frame is equipped with a glass plate, which can be made into a gel plate with a thickness of 1mm and 1.5mm, and a glass plate and a comb with an appropriate thickness can be selected for use according to needs during operation.
(3) The sample hole mold (comb) is used for electrophoretic sample addition.
(4) 4 pairs of screw rods and nuts for fixing the liquid storage tank, 5 pairs for 28D and 30 models.
(5) Five rubber tubes. Two long ones are used to connect the inlet and outlet of cooling water; two of medium length are used to connect the electrode; the outlet of the middle liquid; the shortest one is used to connect the cooling water between the storage tanks.
(6) One pay for wire.
2. How to use
(1) Cleaning the components The above components must be thoroughly cleaned before assembly. In particular, glass plates and concave grooved rubber frames must be thoroughly cleaned with a foam sponge dipped in a little soap powder or detergent. The cleaned glass plate should be placed on the glass plate holder to dry before use.
(2) Assemble the electrophoresis tank
A,
1. Insert four fixed screws into the corresponding holes of a liquid storage frame (half upper tank), and then put the liquid storage frame on the table.
2. Hold the concave groove rubber mold frame in the left hand, and insert the two sides of the glass plate into the rubber mold frame with the thumb and middle finger of the right hand (note that the fingers cannot touch the glass plate on the glued surface).
3. Place the rubber mold frame with glass on the frame of the liquid storage tank placed upside down, and its lower edge must be aligned with the lower edge of the liquid storage tank frame.
4. Hold the other liquid storage tank frame with both hands and fit the liquid storage tank frame on the table, and make the protruding line on the rubber frame located in the center of the organic glass frame of the liquid storage tank.
5. Install four nuts and tighten the screws. Note that the force should be even when tightening the screws, it is best to use both hands to gradually tighten the double angle position.
6. Put the electrophoresis tank upright and put it on the table. The liquid storage tank with short glass plate is called the upper tank (cathode end), and the liquid storage tank with long glass plate is called the lower tank (anode end). There is a gap between the board and the rubber frame. This gap can be filled with the melted 10% agar along the lower end of the long glass plate. In order to prevent air bubbles from remaining, the air bubbles can be removed by raising the end slightly. The gel can only be poured after the agar has completely solidified.
B,
It is also possible to erect the tank vertically, place the rubber mold frame with glass directly into the tank, tighten the screws symmetrically, and seal the bottom edge with agar.
(3) Pouring glue
1. Fill the separation glue first, and then concentrate the glue after solidification. Connect the cooling water before pouring the glue, and clamp the two rubber tubes connected to the upper and lower liquid storage tanks.
2. Use a thin-tip dropper to suck up the glue and add glue from one end along the gap between the long and short glass plates. In order to prevent the generation of air bubbles, you can pick up the other end a little (the air bubbles go high) and add glue continuously. If the single-layer glue (separation glue) is added, the amount of glue added is about 0.3 cm from the top of the short glass plate. Gently add a little water and gently insert the comb with both hands. After gelation, a concave sample hole can be formed.
3. In order to prevent glue leakage, add distilled water to the upper and lower liquid storage tanks immediately, but the distilled water should not exceed the edge of the short glass plate.
4. After the gel solidifies, you can see a bright area with different refractive index between the sample slot comb and the gel plate.
(4) Add sample
1. Loosen the clips on the two rubber tubes connected to the upper and lower liquid storage tanks, let go of the distilled water in the tank, and then clamp the clips after the water flow is completed.
2. Gently remove the sample comb with both hands, you can see the sample hole with clear boundaries, and absorb the water in the hole (be careful not to touch the literal surface of the hole).
3. In the upper and lower liquid storage tanks, add slow; medium liquid, the liquid height should exceed the upper edge of the short glass plate of the upper liquid storage tank.
4. Use a micro-syringe to draw a certain amount of sample and add it to the concave gel sample hole between the long and short glass plates (note that the sample addition action should be light and the needle cannot touch the glue surface).
(5) The electrophoresis-reservoir wire is connected to the negative electrode of the electrophoresis instrument, and the lower liquid-storage wire is connected to the positive electrode of the electrophoresis instrument. Check the line is correct before electrophoresis according to the required voltage and current.
(6) Remove the rubber plate and run the electrophoresis, loosen the nut, remove the plastic frame, peel off the glass plate, and gently pry with the back of the blade in the gap between the lower corners of the two glass plates to separate the rubber surface from the glass plate. Place the glass plate with the rubber plate on the table, gently lift the film along the bottom of the gel with both hands, and place it in a large Petri dish for dyeing. After the rubber plate is rinsed and decolorized, a clear separation band can be seen.
Note: DYCZ-28B, 29 type sandwich electrophoresis tank use method is the same. DYCZ—28C, 28D and 30 electrophoresis tanks are also used in the same way. Because two rubber plates are made, three liquid storage tanks are actually formed. Please note that the short glass plates of the two glue chambers should be inward during operation, with the negative electrode in the middle and the positive electrode on both sides.
note:
It can be used as an electrophoresis tank for two pieces of glue. When making a piece of glue, all three wires should be inserted into the electrophoresis tank to avoid accidental touching on the table.
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