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Electrophoresis: Electrophoresis is an indispensable analytical tool for molecular biology research. Electrophoresis is generally divided into free interface electrophoresis and zone electrophoresis two major categories, free interface electrophoresis without support, such as isoelectric focusing, isotachophoresis, density gradient electrophoresis and microscopic electrophoresis, this type of electrophoresis is rarely used . 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 an electric field. Because different substances have different charges and molecular weights, they move at different speeds in the electric field. 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. Electrophoresis meter is designed and manufactured based on the above principle. The following briefly describes its use and precautions.
â— How to use
1. First connect the two electrodes of the electrophoresis tank with the DC output of the electrophoresis device with a wire, and be careful not to reverse the polarity.
2. The power switch of the electrophoresis system is turned to the off position, the voltage knob is turned to the minimum, and the voltage stabilizing mode and voltage current range are selected according to the working requirements.
3. Turn on the power and slowly rotate the voltage adjustment knob until the desired voltage is reached. Set the electrophoresis termination time and the electrophoresis will begin.
4. After the work is completed, the knobs and switches should be turned to zero or closed, and the electrophoresis plug should be dialed out.
Precautions
1. After the electrophoresis device is powered on, it is forbidden to touch the electrodes, the electrophoretic material and other possible live parts, and it is also not possible to take and place things in the electrophoresis tank. If it is necessary, it should be powered off first to avoid electric shock. At the same time, the instrument must have a good ground to prevent leakage.
2. After the instrument is powered on, do not temporarily add or remove the output wire plug to prevent short circuit. Although there is a fuse inside the instrument, the short circuit may still cause damage to the instrument.
3. Because the resistance values ​​of different media supports are different, the amount of current passed during electrophoresis is also different, and the swimming speed and the time required for swimming to the end point are also different, so the 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 multiple slots, but care must be taken not to overload it, otherwise it will easily affect the life of the instrument.
5. Under certain special circumstances, it is necessary to check the electrophoresis input status of the instrument and allow it to be switched on without load in the steady state. However, in the steady flow state, the load must be connected and then turned on again, otherwise the voltmeter hands will greatly jump, which will easily cause unnecessary Man for machine damage.
6. Abnormal phenomena, such as loud noises, discharges or abnormal odors, must be cut off immediately during the use to repair and prevent accidents.
One, component parts
(i) A pair of platinum tanks equipped with platinum wire, equipped with a cooling device and electrophoresis buffer outlet.
(2) The concave rubber frame is equipped with a glass plate, and a gel plate with a thickness of 1 mm and 1.5 mm can be manufactured respectively. When necessary, an appropriate thickness of the glass plate and a comb are used as required for operation.
(C) Sample hole mold (comb) for electrophoresis sample loading.
(4) Four pairs of screw rods and nuts for fixing liquid reservoirs, and five pairs of 28D and 30 types.
(v) Five rubber tubes. Two long inlets and outlets for connecting the cooling water; two medium lengths for connecting the electrodes; a middle liquid outlet; and the shortest one for connecting the cooling water between the tanks.
(six) wire 1 pay.
â— Second, how to use
(A) Washing Parts Thoroughly clean the above components before they are assembled. In particular, the glass plate and the concave grooved rubber frame must be thoroughly cleaned with a foam sponge dipped in soap powder or detergent. The cleaned glass plate should be placed on a glass plate holder and allowed to dry before it can be used.
(b) Assembly of the electrophoresis tank
A. 1. Insert the four fixed screws into the corresponding holes in one liquid storage box (half-slot), and then place the liquid storage box on the table.
2. Hold the concave groove rubber frame on the left hand side, and insert the right edge of the thumb and the middle finger to the edge of the glass plate and insert it into the rubber frame (note that the finger cannot touch the glass plate on the filling surface).
3. Place the rubber molded frame with glass on the sink frame, and the lower edge must align with the lower edge of the reservoir frame.
4. Hold the other sump frame with your hands and the sump frame resting on the table with your hands, and place the protruding line on the rubber frame in the center of the plexiglass frame.
5. Install four nuts and tighten the screws. Pay attention to tighten the screws when the force should be uniform, it is best to use both hands gradually tighten the oblique position.
6. The electrophoresis tank is vertically erected and placed on the table. The tank with a short glass plate is called the upper tank (cathode end), and the tank with the long glass plate is called the lower tank (the anode end), in the long glass There is a gap between the plate and the rubber frame. This gap can be filled with molten 10% agar along the lower end of the long glass plate. To prevent trapped air bubbles, one can lift the end slightly to remove bubbles. After the agar has completely solidified, the glue can be poured.
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.
(III) Filling glue
1, the first irrigation separation of glue, to be solidified before filling the gel. Connect the cooling water before filling the rubber and clamp the two rubber tubes connecting the upper and lower reservoirs.
2. Use a fine dropper to suck the glue and add glue from one end along the gap between the long and short glass plates. To prevent air bubbles, pick up the other end slightly (bubbles go high) and continue to add glue. If a single layer of glue (separation of plastic) is to add about 0.3 centimeters from the short end of the glass plate, gently add a little water, gently insert the comb into the hands, and after the gel will be able to form a concave sample hole.
3. In order to prevent the leakage of glue, distilled water should be immediately added to the upper and lower sumps, but distilled water should not exceed the edge of the short glass plate.
4. After the gel has solidified, you can see a bright area with a different refractive index between the sample comb and the gel plate.
(D) Add sample
1. Loosen the clamps on the two rubber pipes that connect the upper and lower reservoirs and let go of the distilled water in the tank. After the water flows, clamp the clamps again.
2, gently remove the sample comb, hands, you can see a clear line of wells, the hole to absorb moisture (note that you must not touch the edge of the hole).
3, in the upper and lower reservoir tank to add slow; medium liquid, the height of the liquid should exceed the upper edge of the upper reservoir short glass.
4. Use a microsyringe to suck a certain amount of sample into the concave gel sample well between the long and short glass plates (note that the sample loading action should be light, and the needle must not touch the plastic surface).
(e) Electrophoresis A reservoir wire is connected to the negative electrode of the electrophoresis apparatus, and the lower reservoir wire is connected to the positive electrode of the electrophoresis apparatus. Check the line is correct, in accordance with the required voltage and current electrophoresis.
(6) Take out the rubber plate and finish electrophoresis. Loosen the nut, remove the plastic frame, and peel off the glass plate. In the lower corner space of the two glass plates, use the back of the blade to gently shake, and separate the glue surface from a glass plate. Place the glass plate with the plastic plate on the table, gently hold the film along the bottom of the gel with both hands, and place it in a large petri dish. After the plastic plate has been rinsed and decolorized, clear bands can be seen.
Note: The same applies to the DYCZ-28B and 29 sandwich electrophoresis tanks. The same applies to the DYCZ-28C, 28D, and 30 electrophoresis tanks. Since two plastic sheets are produced, three reservoirs are actually formed. Please note that during the operation, both short glass plates of the plastic chamber should be inward, the middle is negative, and the two sides are positive.
note. Can be used as two pieces of gel electrophoresis tank, when making a piece of plastic, three leads should be inserted in the electrophoresis tank, so as not to touch the table.