CN102824854A - Electrophoresis apparatus and its application - Google Patents

Abstract

The invention provides a macromolecule separation technology and its application. Specifically, an electrophoresis tank and an electrophoresis system comprising the same are provided. The electrophoresis tank provided by the present invention includes at least two fixedly separated or operable separated electrophoresis regions. Without affecting the normal electrophoresis behavior of macromolecules, These separations allow solutions and macromolecules within different electrophoretic regions to be collected separately. The present invention further provides the application of the electrophoresis tank and the electrophoresis system in the separation, electroelution and concentration of nucleic acid, protein, carbohydrate and virus particles.

Description

An electrophoresis device and its application

technical field

The invention relates to a macromolecular separation technology, in particular to a separation device for macromolecular separation, electroelution and concentration and its application.

Background technique

Electrophoresis is the movement of charged particles or molecules in an electric field. In 1809, the Russian physicist PeHce first discovered the phenomenon of electrophoresis, but it was not until 1937 that the Swedish scientist Arne Tiselius assembled the world's first interface electrophoresis device. Subsequently, electrophoresis technology has been widely used, and then a variety of electrophoresis technologies based on different carriers have been developed. Combined with the use of staining reagents such as silver ammonia staining, Coomassie brilliant blue, etc., the coloring and resolution capabilities of biological samples have been greatly improved, and the application of immune technology has improved the resolution to trace and ultra-trace levels, promoting the application of electrophoresis technology in analytical chemistry. , biochemistry, clinical chemistry, toxicology, pharmacology, immunology, microbiology, food chemistry and many other disciplines and different fields.

Under electric field conditions, negatively charged molecules move toward the anode, and positively charged molecules move toward the cathode. Under the action of unit electric field strength, the distance that charged particles move in unit time (ie mobility) is a constant, which reflects the physical and chemical characteristics of the charged particles. Different charged particles are separated from each other due to different moving distances after a period of electrophoresis in the same electric field due to their different charges, or the same charge but different charge-to-mass ratios. Proportional to time. This technique of separating substances by means of electrophoresis is called electrophoresis. Most biological macromolecules such as proteins, nucleic acids, and polysaccharides have cationic and anionic groups, which can be separated by electrophoresis.

The instruments required for electrophoresis mainly include power supply and electrophoresis tank. To make charged biomacromolecules swim in an electric field, an external electric field must be applied, and the resolution and speed of electrophoresis are closely related to the electrical parameters during electrophoresis. The electrophoresis tank is the core part of the electrophoresis system. According to the principle of electrophoresis, the electrophoresis support is placed between the two electrode buffers, and the electric field connects the two buffers through the electrophoresis support. Different electrophoresis methods use different electrophoresis tanks.

The electrophoresis techniques currently used are mainly divided into several types such as mobile interface electrophoresis, isopoint focusing electrophoresis and zone electrophoresis. Moving interface electrophoresis is to place the ion mixture to be separated at one end of the electrophoresis tank. After the electrophoresis starts, the charged particles move to the other pole. The ions with the fastest swimming speed go at the front, and the other ions follow the electrophoresis speed. Arranged in order to form different zones. Isoelectric focusing electrophoresis is to add the ampholyte to the electrophoresis tank filled with pH gradient buffer. When it is in the buffer solution lower than its own isoelectric point, it will be positively charged and move to the negative electrode; if it is at a high In the buffer solution with its own isoelectric point, it is negatively charged and moves to the positive electrode. When these substances swim into their own unique isoelectric point buffer, their net charges become zero, and then they stop swimming, so that substances with different isoelectric points finally focus on their respective isoelectric point positions, forming a Clear zones with very high resolution.

Zone electrophoresis is the most commonly used electrophoretic technique in biomedical research. In a homogeneous carrier electrolyte, the sample is added to a certain support; under the action of an electric field, the positively or negatively charged ions in the sample move to the negative or positive electrode at different speeds, and are separated from each other to form separate zones. . According to different physical properties of supports, zone electrophoresis can be divided into 1) filter paper electrophoresis; 2) powder electrophoresis: such as cellulose powder, starch, glass powder electrophoresis; 3) gel electrophoresis: such as agar, agarose, silica gel, starch gel, polyacrylamide gel electrophoresis; 4) edge electrophoresis: such as nylon silk and rayon electrophoresis. According to the device type of the support, the zone electrophoresis can be divided into 1) flat plate electrophoresis: the support is placed horizontally, which is the most commonly used electrophoresis method; 2) vertical plate electrophoresis: polyacrylamide gel can be made into vertical plate electrophoresis; 3) Columnar (tubular) electrophoresis: polyacrylamide gel can be poured into an appropriate electrophoresis tube to make tubular electrophoresis. According to the continuity of pH, zone electrophoresis can be divided into 1) continuous pH electrophoresis: such as paper electrophoresis and cellulose acetate film electrophoresis; 2) discontinuous pH electrophoresis: such as polyacrylamide gel disc electrophoresis.

Zone electrophoresis has a wide range of applications in biomedical research. Taking protein research as an example, 1) Polyacrylamide gel electrophoresis can be used to identify protein purity. Polyacrylamide gel electrophoresis has both charge effect and molecular sieve effect. It can separate substances with the same molecular size but different numbers of charges, and can also separate substances with the same number of charges but different molecular sizes. Much higher than the general chromatography and electrophoresis methods, and good repeatability, no electroosmosis. 2) SDS polyacrylamide gel electrophoresis can be used to determine protein molecular weight. The principle is that SDS with a large amount of charge is bound to the protein molecule, which overcomes the influence of the original charge of the protein molecule and obtains a constant charge/mass ratio. The measurement time is short, the resolution is high, and only a very small amount of sample (1-100ug ). 3) Polyacrylamide gel electrophoresis can be used for protein quantification. The gel after electrophoresis is scanned by a gel scanner to give quantitative results. 4) Agar or agarose gel immunoelectrophoresis can be used to check the purity of protein preparations, analyze the components of protein mixtures, study whether antiserum preparations have antibodies against a known antigen, and test whether two antigens are the same, etc.

In biological experiments, it is often necessary to recover the macromolecular substances separated by electrophoresis from the electrophoresis carrier (agarose gel or polyacrylamide gel), so that further analysis can be performed. In this process, the most critical considerations include the quality (purity and concentration) of the recovered product, recovery efficiency, and ease of operation. 1) The quality of recovered products is one of the most critical technical indicators. During routine recovery, common grade agarose with some unidentified polysaccharides will be extracted from the gel along with the DNA. These unknown substances may strongly inhibit subsequent ligation, enzyme digestion, or labeling, amplification and other reactions. For large fragment DNA recovery, the quality of the recovered product also includes the integrity of the DNA fragment, while for smaller fragments, the concentration of the recovered product is an important consideration. 2) Recovery yield is another important parameter. Since the amount of electrophoresis sample is usually very small, the electrophoresis process itself will also lead to the dispersion and loss of the sample. Therefore, it is important for subsequent research to recover as many target fragments as possible in the electrophoresis gel bands and improve the product yield. very important. 3) The convenience of operation is another key factor, because gel recovery is a basic experimental operation, and a simple, fast and low-cost research method needs to be established.

Among the various gel recovery operations applied in the laboratory, the simplest method is to cut off the gel block containing the separated macromolecules, and to make the separated macromolecules from the gel by mechanical fragmentation and long-term buffer elution. spread out.

Low-melting point agarose gels are another, simpler gel recovery technique. Prepare the gel with low melting point agarose, cut the target band after electrophoresis, keep warm in TE solution to melt the gel, extract with traditional phenol chloroform, and precipitate with ethanol. This method requires the use of organic reagents such as phenol chloroform and takes a long time.

The glass milk/purified filler gel recycling method is another more flexible recycling method. First cut off the strips of the electrophoresis gel, dissolve the gel in the provided solution, add purification fillers to absorb and mix, quickly centrifuge to remove the supernatant, wash the precipitate, and use the eluent to purify the nucleic acid fragments adsorbed in the medium. This method is suitable for fragments of various sizes, especially the recovery of large fragments, but the operation is more complicated than the former, involving multiple centrifugation and supernatant collection.

The DEAE cellulose film paper method and its improved method are also a relatively traditional glue recovery technology. First cut the DEAE cellulose membrane into small strips for activation treatment; after electrophoresis for a period of time, cut a knife in front of the target band, insert a DEAE cellulose membrane slightly wider than the band into the cut, and continue electrophoresis for a while, the DNA on the band Retained by the diaphragm, take out the diaphragm and wash it, then transfer it to a centrifuge tube and add buffer to keep warm for elution, then perform phenol-chloroform extraction and ethanol precipitation. Obviously, this manual recovery method has high technical requirements for experimenters, poor repeatability, and is not suitable for large-scale operations.

In addition to these more traditional methods, electroelution techniques are now widely used in the field of biological research. Electroelution refers to the technique of migrating out the target components contained in certain supports by electrophoresis. The specific method is to place the gel containing the separated macromolecules in a space separated by a semi-permeable membrane, and the electrophoresis current causes the DNA to leave the gel and enter the liquid phase, and the liquid phase is recovered to purify the DNA molecules therein. Using electroelution technology, a series of convenient and applicable electroelution devices have been developed in recent years, including US Pat. US Pat. No. 4,964961 and US Pat. No. 5,340,449. The electroelution condition is mild, the operation is simple, and there is no special requirement for agarose, and it can also be used for fragment recovery or protein recovery in acrylamide gel; compared with other traditional gel recovery techniques, it has great advantages . However, due to the selective permeation properties of semi-permeable membranes, it is necessary to select a suitable semi-permeable membrane according to the size of the separated molecules, which limits the application of this technology; in addition, the adsorption of semi-permeable membranes to recovered products is also A problem that cannot be ignored.

Contents of the invention

The present invention relates to the subject matter defined in the following sequentially numbered paragraphs:

1. An electrophoresis tank, characterized in that the electrophoresis tank comprises at least two fixedly separated or operable separated electrophoresis regions, and these separations make the different electrophoresis regions Solutions can be collected separately.

2. An electrophoresis tank, characterized in that the electrophoresis tank includes two fixedly separated or operable separated electrophoresis regions, and these separations make the solutions in different electrophoresis regions can be collected separately.

3. An electrophoresis tank, characterized in that the electrophoresis tank includes three fixedly separated or operable separated electrophoresis regions, and these separations make the solutions in different electrophoresis regions can be collected separately.

4. The electrophoresis tank according to paragraphs 1-3, characterized in that the materials used to fix and separate different electrophoresis regions include agarose gel, polyacrylamide gel, and solid phase supports with holes.

5. The electrophoresis tank according to paragraphs 1-3, wherein the means for operable separation of different electrophoresis areas include valves, switches, and blockable flow channels.

6. The electrophoresis tank according to paragraph 1, characterized in that the electrophoresis tank comprises a tank body, an anode, a cathode and an electrophoresis area; the electrophoresis area is composed of three areas separated by agarose gel or polyacrylamide gel, and the anode and cathodes are respectively located in the regions at both ends.

7. The electrophoresis tank according to paragraph 1, characterized in that the electrophoresis tank includes a tank body, an anode, a cathode and an electrophoresis area; the electrophoresis area includes three independent areas, and the areas are connected through an electrophoresis channel, and the anode and the cathode are respectively located at In separate regions at both ends; electrophoresis channel with operable separators.

8. The electrophoresis tank according to paragraph 1, characterized in that the electrophoresis tank comprises a tank body, an anode, a cathode and an electrophoresis area; wherein, the electrophoresis area consists of two nested separable cylindrical or tubular structures and between them composed of connecting channels.

9. An electrophoresis system, characterized in that the electrophoresis system includes a power supply and the electrophoresis tank described in any one of paragraphs 1-8.

10. Use of the electrophoresis system described in paragraph 9 in the separation, electroelution and concentration of macromolecules.

11. The use according to paragraph 10, characterized in that the macromolecules include nucleic acid molecules, protein molecules, carbohydrate molecules, and virus particles.

12. Use of the electrophoresis system described in paragraph 9 in the separation of nucleic acids and proteins.

The technical problem to be solved by the present invention is how to separate macromolecular substances efficiently, conveniently and at low cost. The most important technical improvement of the present invention is to distribute macromolecules with different charge properties in different electrophoretic regions that are separated or operable by electrophoresis, and separate different macromolecules by separately collecting solutions in different electrophoretic regions .

In one aspect, the present invention provides an electrophoresis tank, the electrophoresis tank includes at least two fixedly separated or operable separated electrophoresis regions, and these separations allow different electrophoresis regions to The solution can be collected separately.

In another aspect, the present invention provides an electrophoresis tank, which includes two fixedly separated or operable separated electrophoretic regions, and these separations allow different electrophoretic regions to The solution can be collected separately.

On the other hand, the present invention provides an electrophoresis tank, which includes three fixedly separated or operable separated electrophoretic regions, and these separations allow different electrophoretic regions to The solution can be collected separately.

According to the present invention, the materials used to fix and separate different electrophoretic regions include but not limited to agarose gel, polyacrylamide gel, solid phase support with holes.

According to the present invention, means for operable separation of different electrophoretic regions include valves, switches, and flow channels that can be blocked.

According to the present invention, the provided electrophoresis tank includes a tank body, an anode, a cathode and an electrophoresis area; the electrophoresis area is composed of three areas separated by agarose gel or polyacrylamide gel, and the anode and the cathode are respectively located in the areas at both ends .

According to the present invention, the provided electrophoresis tank includes a tank body, an anode, a cathode, and an electrophoresis area; the electrophoresis area includes three independent areas, and the areas are connected through an electrophoresis channel, and the anode and the cathode are respectively located in independent areas at both ends; the electrophoresis channel With operable divider.

According to the present invention, the provided electrophoresis tank includes a tank body, an anode, a cathode and an electrophoresis area; wherein, the electrophoresis area is composed of two nested separable cylindrical or tubular structures and a connecting channel between them.

On the other hand, the present invention provides an electrophoresis system, which includes a power supply and the electrophoresis tank provided by the present invention.

In another aspect, the present invention provides the application of the electrophoresis system in the separation, electroelution and concentration of macromolecules.

According to the present invention, the macromolecules include nucleic acid molecules, protein molecules, carbohydrate molecules and virus particles.

According to the present invention, the application of the electrophoresis system in the separation of nucleic acid and protein.

Beneficial effects of the present invention

In the present invention, the electrophoresis tank is divided into several electrophoresis regions by using fixed partitions or operable partitions, so that different macromolecules can be distributed in different regions under the action of an electric field according to their own charge properties without affecting the normal electrophoresis behavior of macromolecules. In the electrophoresis area, the macromolecular substances existing in each area are separated by collecting the solutions in different areas. Isolated macromolecules of the invention include, but are not limited to, nucleic acids, proteins, carbohydrates, and viral particles. Compared with the prior art, the separation device and technology provided by the invention can separate macromolecular substances more simply, efficiently and at low cost.

Description of drawings

Figure 1. A schematic diagram of a two-chamber electrophoresis tank separated by agarose

Figure 2. A schematic diagram of a three-chamber electrophoresis tank separated by agarose

Figure 3. A schematic diagram of a separate two-chamber electrophoresis tank

Figure 4. Schematic diagram of a separate three-chamber electrophoresis tank

Figure 5. Schematic diagram of a casing electrophoresis tank

Figure 6. Schematic diagram of a casing electrophoresis tank

Detailed ways

The electrophoresis system provided by the present invention includes a power source and an electrophoresis tank, and the electrophoresis tank includes a tank body, an anode, a cathode, and an electrophoresis area between the anode and the cathode. Among them, the electrophoretic region is divided into at least two regions by fixed or operable partitions. These fixed or temporary partitions can make the solutions in each region and the macromolecular samples in them It can be collected separately, so as to realize the separation and recovery of macromolecular substances. The body of the electrophoresis tank may be fabricated from suitable materials including, but not limited to, glass, plexiglass, plastic, resin, polypropylene, acrylic, or similar materials. The body of the electrophoresis tank can be in any suitable shape, including but not limited to square, rectangular, triangular, circular, cylindrical, spherical, conical or a combination of different shapes. The separated electrophoresis area can be divided into anode area, cathode area and intermediate area according to the relationship with the electrodes; the volume of each area can be freely set at 10uL or more to adapt to different uses of the electrophoresis tank.

According to the present invention, the material for fixing and separating the electrophoresis tank into different regions can be selected from agarose gel, polyacrylamide gel, and a solid phase support with holes or micropores; the solid phase support can include dialysis Other suitable materials other than membranes, semipermeable membranes, filters, and filter papers include, but are not limited to, plexiglass, plastic, resin, polypropylene, acrylic, or similar materials.

According to the present invention, the means for operably separating the electrophoresis tank into different areas may include valves, switches, blockable flow channels, or disassembly and separation of different electrophoresis areas. The blockable channel means that the channel connecting different electrophoretic regions can be physically blocked to achieve operable separation. The specific blocking methods include but are not limited to clamping with hemostats, folding the channel to achieve separation, Insert barriers to achieve separation.

The present invention will be described in further detail below in conjunction with examples. It should be understood that these examples are listed for illustrative purposes only, and are not intended to limit the scope of the present invention. Unless otherwise specified, the reagents and culture media used in the present invention are commercially available. The experimental method that does not indicate specific condition in the embodiment, usually carries out according to conventional experimental condition, such as Sambrook et al. in " Molecular Cloning: Laboratory Handbook " (New York: Cold Spring Harbor Laboratory Press, 1989) described condition, or Follow the conditions recommended by the manufacturer.

Embodiment 1. Electrophoresis tank structure

An agarose-separated two-chamber electrophoresis tank

Referring to Fig. 1, the electrophoresis tank comprises a tank body 1, an anode 2, a cathode 3, an anode area 4, an intermediate area 5 and an agarose gel partition 7; An anode region 4 and an intermediate region 5, respectively, in which are located the anode 2 and the cathode 3, respectively.

A kind of agarose separated three-chamber electrophoresis tank

Referring to Fig. 2, this electrophoresis tank comprises tank body 1, anode 2, cathode 3, anode region 4, middle region 5, cathode region 6 and agarose gel partition 7; The anode 2 and the cathode 3 are respectively located in the anode region 4 and the cathode region 6 at both ends.

A separate two-chamber electrophoresis tank

Referring to Fig. 3, the electrophoresis tank includes a tank body 1, an anode 2, a cathode 3, an anode area 4, an intermediate area 5, a valve 8 and a connecting channel 9; the electrophoresis area includes two independent areas, namely the anode area 4 and the intermediate area 5, they are connected by an electrophoresis channel 9, the anode 2 and the cathode 3 are respectively located in the anode area 4 and the middle area 5; the electrophoresis channel 9 has a valve 8.

A separate three-chamber electrophoresis tank

Referring to Fig. 4, the electrophoresis tank includes a tank body 1, an anode 2, a cathode 3, an anode area 4, an intermediate area 5, a cathode area 6, a valve 8 and a connecting channel 9; the electrophoresis area includes three independent areas, namely the anode area 4. The middle area 5 and the cathode area 6 are connected by the electrophoresis channel 9, the anode 2 and the cathode 3 are respectively located in the anode area 4 and the cathode area 6; the electrophoresis channel 9 has a valve 8.

A casing type electrophoresis tank

Referring to Fig. 5, the electrophoresis tank includes a tank body 1, an anode 2, a cathode 3, an anode area 4, an intermediate area 5 and a connecting channel 10; the electrophoresis area consists of two nested separable tubular structures and a connecting channel between them 10 composition; the anode area 4 is located in the outer tube, the middle area 5 is located in the inner tube, the anode 2 and the cathode 3 are located in the anode area 4 and the middle area 5 respectively.

A casing type electrophoresis tank

Referring to Fig. 6, the electrophoresis tank includes a tank body 1, an anode 2, a cathode 3, an anode area 4, an intermediate area 5 and a connecting channel 10; the electrophoresis area consists of two nested separable tubular structures and a connecting channel between them 10 composition; the anode area 4 is located in the inner tube, the middle area 5 is located in the outer tube, the anode 2 and the cathode 3 are respectively located in the anode area 4 and the middle area 5.

Example 2. Electroelution of Nucleic Acid Molecules in Gels

The plasmid band separated in the agarose gel was recovered with the sleeve type electrophoresis tank shown in Figure 6 .

1. Plasmid electrophoresis: take 20uL (4ug) of the purified pGL3 (Promega) plasmid, mix it with 2uL loading buffer, load it on a 1.2% agarose gel, and perform electrophoresis under constant voltage conditions of 100V. Liquid is 0.5×TBE;

2. When the plasmid band moves to the middle of the agarose gel, stop the electrophoresis, take out the gel block, stain with ethidium bromide, and cut off the gel strip containing the plasmid band with a clean blade under ultraviolet light ;

3. Put the gel strip into the outer tube of the electrophoresis tank shown in Figure 6, add 400uL 0.5×TBE to immerse the gel strip; add 100uL 0.5×TBE into the inner tube, so that the connecting channel of the inner and outer tubes is filled with electrophoresis buffer liquid, forming a current loop between the inner and outer tube solutions;

4. Connect the power electrodes as shown in Figure 6, and after 10 minutes of electrophoresis at a constant voltage of 10V, stop the electrophoresis and separate the inner and outer tubes;

5. Precipitate the plasmid in the inner tube solution with ethanol, dissolve the precipitate with 10uL deionized water, detect the purity of the plasmid and quantify it, and calculate the recovery rate;

6. A260/280 is 1.91, and the recovery rate is 94%.

Experimental results show that the sleeve-type electrophoresis tank provided by the present invention can recover nucleic acid bands separated in gel electrophoresis in a simple, efficient and high-quality manner.

Example 3. Electroelution of protein molecules in gels

Use the sleeve type electrophoresis tank shown in Figure 6 to recover the protein bands separated in the PAGE gel.

1. Protein electrophoresis: take 10ug bovine serum albumin (Sigma), mix it with loading buffer and load it on a 10% PAGE gel, the electrophoresis condition is 200v/10mA, and the electrophoresis buffer is Tris-glycine protein electrophoresis buffer liquid;

2. When the loading indicator swims to the 2/3 position of the gel, stop the electrophoresis, take out the gel block, and cut off the gel strip containing the target protein band with a clean blade;

3. Put the gel strip into the outer tube of the electrophoresis tank shown in Figure 6, add 400uL 50mM Tris (pH8.3) solution to immerse the gel strip; add 100uL 50mM Tris (pH8.3) solution to the inner tube to make the connection The connecting channel of the inner and outer tubes is filled with Tris buffer, forming a current loop between the inner and outer tube solutions;

4. Connect the power electrodes as shown in Figure 6. After electrophoresis at a constant voltage of 15V for 20 minutes, stop the electrophoresis and separate the inner and outer tubes;

5. Use the Bradford method to measure the protein content in the inner tube solution, and calculate the recovery rate;

6. The calculated recovery rate is 93%.

Experimental results show that the sleeve-type electrophoresis tank provided by the invention can simply and efficiently recover protein bands separated in gel electrophoresis.

Example 4. Nucleic acid enrichment

The nucleic acid components in the solution were concentrated using a two-chamber electrophoresis tank separated by agarose gel as shown in FIG. 1 . The volume of the anode area of the electrophoresis tank is 20mL, the volume of the middle area is 200mL, the width of the agarose strip is 1cm, and the top surface exceeds the liquid level height of the electrophoresis buffer by 0.5cm, forming a separation between the anode area and the middle area of the electrophoresis tank .

1. Take 20ug of purified pGL3 (Promega) plasmid, dissolve it in 200mL0.5×TBE, and the final concentration of the plasmid is 100ng/mL;

2. Slowly add 200mL of plasmid solution to the middle area of the electrophoresis tank, and add 20mL of 0.5×TBE to the anode area of the electrophoresis tank;

3. Connect the power supply electrodes as shown in Figure 1, and stop the electrophoresis after electrophoresis at a constant voltage of 40V for 20 minutes;

5. Collect the solution in the anode area of the electrophoresis tank, carry out ethanol precipitation, dissolve the precipitate with 15uL deionized water, detect the purity of the plasmid and quantify it, and calculate the recovery rate;

6. A260/280 is 1.87, and the recovery rate is 95%.

Experimental results show that the partitioned electrophoresis tank provided by the invention can concentrate nucleic acid molecules in samples simply, efficiently and with high quality.

Example 5. Separation of Nucleic Acid and Protein

Use the separate two-chamber electrophoresis tank shown in Figure 3 to separate nucleic acid and protein molecules from the nucleic acid-protein mixture. The volume of the anode area of the electrophoresis tank is 20mL, the volume of the middle area is 200mL, and a valve is installed in the electrophoresis channel connecting the anode area and the middle area.

1. Nucleic acid protein mixture: Take 10ug of purified pGL3 (Promega) plasmid and 10mg of bovine serum albumin (Sigma), dissolve in 200mL0.5×TBE, the final concentration of plasmid is 50ng/mL, and the final concentration of protein is 50ug/mL;

2. Close the valve in the electrophoresis channel, add 200mL nucleic acid protein solution to the middle area of the electrophoresis tank, and add 20mL0.5×TBE to the anode area of the electrophoresis tank;

3. Connect the power electrodes as shown in Figure 3, open the valve, and stop the electrophoresis after 20 minutes of electrophoresis at a constant voltage of 40V;

5. Collect the solution in the anode area of the electrophoresis tank, carry out ethanol precipitation, dissolve the precipitate with 15uL deionized water and quantify it, and calculate the recovery rate; use the Bradford method to measure the protein content in the solution in the anode area and the middle area respectively;

6. The recovery rate of plasmid DNA was 97%; the protein concentration in the anode region was 5ug/mL, and the protein concentration in the middle region was 47ug/mL.

Experimental results show that the separation type electrophoresis tank provided by the invention can separate nucleic acid and protein molecules from nucleic acid protein mixture in a simple, efficient and high-quality manner.

Claims (10)

1. electrophoresis tank; It is characterized in that said electrophoresis tank comprises that at least two fixing electrophoresis that separate or that operability is separated are regional; Under the situation that does not influence the normal electrophoresis behavior of big molecule, these separations make the solution in the different electrophoresis zone can be by independent collection.

2. electrophoresis tank is characterized in that said electrophoresis tank comprises two fixing separate or electrophoresis zones that operability is separated, and under the situation that does not influence the normal electrophoresis behavior of big molecule, these separations make the solution in the different electrophoresis zone can be by independent collection.

3. electrophoresis tank is characterized in that said electrophoresis tank comprises three fixing separate or electrophoresis zones that operability is separated, and under the situation that does not influence the normal electrophoresis behavior of big molecule, these separations make the solution in the different electrophoresis zone can be by independent collection.

4. according to the described electrophoresis tank of claim 1-3, it is characterized in that being used for fixing the material of separating different electrophoresis zone and comprise Ago-Gel, polyacrylamide gel, solid support with holes.

5. according to the described electrophoresis tank of claim 1-3, it is characterized in that being used for operability and separate the mode in different electrophoresis zone and comprise valve, switch, property runner capable of blocking.

6. electrophoresis tank according to claim 1 is characterized in that this electrophoresis tank comprises cell body, anode, negative electrode and electrophoresis zone; The electrophoresis zone is by being formed by three zones of Ago-Gel or polyacrylamide gel separation, and anode and negative electrode lay respectively in the zone at two ends.

7. electrophoresis tank according to claim 1 is characterized in that this electrophoresis tank comprises cell body, anode, negative electrode and electrophoresis zone; The electrophoresis zone comprises three distinct area, and said zone connects through electrophoresis path, and anode and negative electrode lay respectively in the isolated area at two ends; Electrophoresis path has exercisable separating device.

8. electrophoresis tank according to claim 1 is characterized in that this electrophoresis tank comprises cell body, anode, negative electrode and electrophoresis zone; Wherein, the electrophoresis zone is made up of two nested separable tubulars or tubular structure and the interface channel between them.

9. an electrophoresis system is characterized in that said electrophoresis system comprises power supply and each described electrophoresis tank of claim 1-8.

10. the described electrophoresis system of claim 9 is in macromolecular separation, electroelution with the application in concentrating.

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