CN106754893A - Electroelution instrument - Google Patents
Electroelution instrument Download PDFInfo
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- CN106754893A CN106754893A CN201710099124.3A CN201710099124A CN106754893A CN 106754893 A CN106754893 A CN 106754893A CN 201710099124 A CN201710099124 A CN 201710099124A CN 106754893 A CN106754893 A CN 106754893A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/24—Extraction; Separation; Purification by electrochemical means
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Abstract
The invention discloses a kind of electroelution instrument, belong to separation and the purification technique field of biological sample, there is provided a kind of simple structure, easy to operate, lower-cost electroelution instrument, including base and electroelution cup;There is electroelution cavity inside the electroelution cup;The electrode slice of sheet is respectively arranged with the relative wall of cup in the electroelution cavity both sides, and the one side of the electrode slice exposes from the outer surface of electroelution cup;It is every to be connected by the conductive mouth through wall of cup between plate electrode piece and electroelution cavity;Slot is provided with the base, electroconductive elastic sheet is respectively arranged with the both sides of the slot.Electroelution instrument of the present invention need to be only put into electroelution cavity sample to be eluted when using, and be subsequently adding appropriate buffer solution;Energization wash-out treatment is carried out by electroelution cup is integrally inserted into corresponding slot afterwards.
Description
Technical field
Separation and purification technique field the present invention relates to biological sample, more particularly to a kind of electroelution instrument.
Background technology
In the operation of molecular biology experiment, to be seen using agarose and polyacrylamide gel electrophoresis in many cases
Examine, purify and reclaim DNA fragmentation and protein.It is important step therein, experimental mesh to reclaim DNA fragmentation and protein
Difference, the DNA of recovery can be used for carrier connection, genetic recombination, sequence analysis, probe and the follow-up work such as prepares;And reclaim
Protein is then usually used in Antibody preparation and structural analysis.So, concentration and purity to reclaiming target have certain requirement.
During practice, accurate, easy, quick, economic way of recycling, is all the time the pursuit of experimenter.
Reclaim DNA fragmentation and protein from Ago-Gel or polyacrylamide gel, it is main using wash-out-collect and
Be coupled-combine two ways.According to the difference of wash-out principle, numerous elution process can be summarized as two classes:Direct elution method
And electroelution method.Direct elution method be DNA fragmentation protein under the immersion of solvent free diffusing and go out, electroelution rule is profit
Diffused out with electrically charged mutually the absorption with electrode of institute on DNA fragmentation and protein.It is through the sample after wash-out then purified, dense
The steps such as contracting so that sample reaches subsequent experimental requirement.
Paces over time, electroelution method is gradually shown up prominently, and bag filter electroelution rule is because it is relatively preferable
Operability and reliability, become the common method of electroelution.Its concrete operations is as follows:After sample electrophoresis are finished, cutting needs
The zone of recovery, gel strips are placed in the bag filter containing appropriate amount of buffer solution, place into electrophoresis 30-60min in electrophoresis tank, are made
On the wall of target dna and protein molecule swimming to bag filter by positive pole side, then counterelectrophoresis 30-60s, makes to be attached to dialysis
DNA and protein molecule swimming on mould are in the buffer solution that PH in bag filter is 8.0.Buffer solution in sucking-off bag, purified,
The steps such as precipitation reclaim target DNA fragment and protein.
Classical processes in traditional direct elution method and coupling-combined techniques --- LMP gel method, in operating process
In use high concentration salt solutions, there is a certain degree of damage to DNA fragmentation and protein, have to subsequent experimental one be fixed
Ring.And the method such as electroelution method, bag filter electrophoresis and DEAE fiber membrane electrophoresis derived by electric charge tractive force, though certain journey
Degree compensate for the deficiency of conventional method, but also because the equipment without matching causes cumbersome and efficiency is not good enough.Nowadays,
MERCK (Merck) company is improved on the basis of bag filter electrophoresis, devises special D-tube electroelution pipes, should
The method that electroelution pipe enhances bag filter electrophoresis really, but asks without solution versatility in improving bag filter electrophoresis completely
Topic, in addition it is also necessary to needed according to subsequent experimental and replace new buffer solution, so whole process is still relatively complicated.
Conventional electroelution device mainly has the D-tube electroelutions pipe and U.S. Bio- of MERCK (Merck) company now
The type electroelution instrument of Bole 422 of Rad (Bole) company.There is much in common in face in both, be all using semi-transparent in design
Film is intercepted to reclaiming fragment, is then that operation can just obtain purpose fragment carrying out one, therefore its operating process is more numerous
It is trivial.In addition, the latter is with respect to for the former, although avoid influence of the buffer solution to subsequent experimental, but need additional extra
Multiple centrifugal process, therefore its operating process is also relatively complicated.
The content of the invention
Present invention solves the technical problem that being to provide a kind of simple structure, easy to operate, lower-cost electroelution instrument.
The technical solution adopted for the present invention to solve the technical problems is:Electroelution instrument, including base and electroelution cup;
There is electroelution cavity inside the electroelution cup;It is respectively arranged with the relative wall of cup in the electroelution cavity both sides
The electrode slice of sheet, and the one side of the electrode slice exposes from the outer surface of electroelution cup;Washed with electricity per plate electrode piece
Connected by the conductive mouth through wall of cup between de- cavity;Slot is provided with the base, in the both sides of the slot point
Electroconductive elastic sheet is not provided with it;The electroelution cup is removably inserted into the slot, and when the electroelution cup
After being inserted into the slot, the electroconductive elastic sheet that the electrode slice can be respectively with respective side electrically conducts.
It is further:The profile of the electroelution cup is in cuboid, and its one end is provided with opening portion;Institute
Electrode slice is stated to be arranged on the relative wall of cup of the one of which of electroelution cup surrounding.
It is further:Electrode slice on wall of cup is in specular and the setting being parallel to each other.
It is further:The one end for being provided with opening portion is the top of electroelution cup, and one end relative with opening portion is
The bottom of electroelution cup;In the bottom of electroelution cup, prolong to middle part inclination after being provided with the both sides wall of cup bending of electrode slice
Stretch, and V-arrangement ditch slot structure or inverted trapezoidal groove structure are formed in the bottom of electroelution cavity.
It is further:The conductive mouth is elongated, and length direction and the electroelution cavity of conductive mouth depth
Direction is consistent.
It is further:Multiple slots independent of each other are provided with base.
It is further:Voltage-regulation knob is additionally provided with base.
The beneficial effects of the invention are as follows:It is simple structure of the present invention, easy to use, sample to be eluted need to only be put when using
Enter in electroelution cavity, be subsequently adding appropriate buffer solution;Electroelution cup is integrally inserted into corresponding slot afterwards
Energization wash-out treatment can be carried out;After electroelution treatment, the buffer solution in electroelution cup also can be very easily processed.And
And, further by setting the bottom of electroelution cavity in groove structure, gel sample can met according to initial gel volume
Add appropriate buffer solution volume on the premise of being totally submerged, so be more convenient for the buffer solution volume that is used during control wash-out and
Buffer solution volume during recovery.In addition, the electroelution cup in the present invention is also reusable, therefore its cost is lower.
Brief description of the drawings
Fig. 1 is the top view of electroelution instrument of the present invention;
Fig. 2 is the three-dimensional view of electroelution instrument of the present invention;
Fig. 3 is the schematic diagram after hiding electroelution cup in Fig. 1;
Fig. 4 is the schematic diagram after hiding electroelution cup in Fig. 2;
Fig. 5 is the three-dimensional view of single electroelution cup;
Fig. 6 is the front view of single electroelution cup;
Fig. 7 is the top view of single electroelution cup;
Fig. 8 is the sectional view of Section A-A in Fig. 7, and the wherein bottom of electroelution cavity is in inverted trapezoidal groove structure;
Fig. 9 is the sectional view of Section A-A in Fig. 7, the V-shaped groove structure in bottom of wherein electroelution cavity;
Figure 10 is the circuit connecting relation schematic diagram inside electroelution instrument in the present invention;
Marked in figure:Base 1, electroelution cup 2, wash-out cavity 3, electrode slice 4, conductive mouth 5, slot 6, electroconductive elastic sheet
7th, opening portion 8, V-arrangement ditch slot structure 9, inverted trapezoidal groove structure 10, voltage-regulation knob 11, power switch 12, power supply terminal 13.
Specific embodiment
The present invention is further described with reference to the accompanying drawings and detailed description.
As shown in Fig. 1 to Figure 10, electroelution instrument of the present invention, including base 1 and electroelution cup 2;Described
The inside of electroelution cup 2 has electroelution cavity 3;Piece is respectively arranged with the relative wall of cup in the both sides of electroelution cavity 3
The electrode slice 4 of shape, and the one side of the electrode slice 4 exposes from the outer surface of electroelution cup 2;Per plate electrode piece 4 with electricity
Connected by the conductive mouth 5 through wall of cup between wash-out cavity 3;Slot 6 is provided with the base 1, in the slot 6
Both sides are respectively arranged with electroconductive elastic sheet 7;The electroelution cup 2 is removably inserted into the slot 6, and when the electricity
After wash-out cup 2 is inserted into the slot 6, the electroconductive elastic sheet 7 that the electrode slice 4 can be respectively with respective side electrically conducts.
Wherein, the effect of electroelution cup 2 is washed with carrying out electricity for containing sample to be eluted and corresponding buffer solution
De- treatment, you can put into electroelution cavity 3 sample to be eluted and add appropriate buffer solution, is then electrified to realize
Electroelution treatment.It is to be removably mounted on base 1 in corresponding slot 6 for electroelution cup 2 in the present invention, and electricity
Wash-out cup 2 may be such that the cost of electroelution instrument of the present invention is lower as reusable part.
More specifically, preferably the profile of the electroelution cup 2 is in cuboid, and its one end is provided with opening
Portion 8;The electrode slice 4 is arranged on the relative wall of cup of the one of which of the surrounding of electroelution cup 2;Specifically can refer to the He of accompanying drawing 5
Shown in accompanying drawing 7.
In addition, it is above-mentioned electroelution cup 2 is set to cuboid when, further can will be located at wall of cup on electrode slice
4 in specular and the setting that is parallel to each other, as shown in Figure 7.
In addition, the effect of conductive mouth 5 is for being connected electrode slice 4 with the electroelution cavity 3 of inside, so being may be such that
When carrying out electroelution operation, the buffer solution being added in electroelution cavity 3 can be contacted directly with electrode slice 4.Specifically, settable
The conductive mouth 5 is elongated, and the length direction of conductive mouth 5 is consistent with the depth direction of electroelution cavity 3;Such as accompanying drawing 5
Shown in 7;And conductive mouth 5 under normal circumstances, is arranged on the medium position on corresponding wall of cup, and both sides will be located at
Conductive mouth 5 is symmetrical set.Width and length as conductive mouth 5 etc., can as the case may be depending on, strict limit
System.
In addition, the purpose that opening portion 8 is set on electroelution cup 2 is for waiting to elute to being put into electroelution cavity 3
Sample and add corresponding buffer solution;Specifically, it has been generally acknowledged that the one end for being provided with opening portion 8 is electroelution cup 2
Top, one end relative with opening portion 8 is the bottom of electroelution cup 2 accordingly;The present invention is further in electroelution cup 2
Bottom, inclines extension, and form V-arrangement in the bottom of electroelution cavity 3 after being provided with the both sides wall of cup bending of electrode slice 4 to middle part
Groove structure 9 or inverted trapezoidal groove structure 10, specifically can refer to shown in accompanying drawing 8 and accompanying drawing 9.By in electroelution cavity
3 bottom sets corresponding groove structure, and electroelution cup 2 can be made at least to have following 3 benefits:First is to carry out electricity
The bottom and the direction on top of electroelution cup 2 can be more conducive to distinguish during elution action, can so be avoided by electroelution
Cup 2 is inserted into maloperation when in slot 6;Second allows for sample to be eluted typically sheet gel structure, therefore
By be arranged to groove structure can avoid sample electroelution cavity 3 bottom occur bending, overlap etc. " on all fours " phenomenon with
And thereby result in wash-out distance and become big problem, and then may be such that after the sample for eluting is put into electroelution cavity 3 and be in
Upright state, it is most short to ensure wash-out distance, and then be more conducive to carry out electroelution treatment to sheet gel sample;3rd be by
It is groove structure in the bottom of electroelution cavity 3, therefore its corresponding cross-sectional area is relative to the horizontal stroke of the non-groove structure of top
Sectional area is smaller, therefore cavity corresponding to same units depth is smaller so that the bottom of electroelution cavity 3 accommodated it is slow
Fliud flushing is less, therefore by controlling the buffer solution of the upper end of electroelution cavity 3 on the premise of being totally submerged gel sample is met
When the buffer solution volume used when liquid level is to control total buffer solution volume, and then control of being more convenient for elute and recovery
Buffer solution volume.
In addition, the present invention can also be provided with multiple slots independent of each other 6 on base 1, as shown in Figure 1 specific
Four slots 6 are provided with diagram.So, in theory can simultaneously to inserting an electroelution cup 2 respectively in four slots 6,
And then can be while the sample to be eluted to four carries out electroelution treatment.Certainly, without loss of generality, can also be set on base 1
There is greater number of slot 6, and when the slot 6 of two and the above is provided with, each slot 6 can be set can be separately
Carry out electroelution treatment, you can optionally carry out electroelution treatment using only one of slot 6.
In addition, the shape and structure of slot 6 should be mutually matched with electroelution cup 2;And should be inserted in electroelution cup 2
After entering in slot 6, can be electrically conducted with the electrode slice 4 of respective side respectively by the electroconductive elastic sheet 7 on the madial wall of slot 6.
Specifically, electroconductive elastic sheet 7 may be configured as the metallic sheet-like formation with certain elasticity, when electroelution cup 2 is inserted into slot 6
When, its electrically conducting between electrode slice 4 is ensured by the elastic reaction of electroconductive elastic sheet 7.Without loss of generality, can also be in slot
6 oral area is provided with corresponding lid (not drawn in figure) or the corresponding lid of optional equipment, and when electroelution cup 2 is inserted
After in slot 6, the opening portion 8 of electroelution cup 2 can be covered by corresponding lid.Common, above-mentioned lid can be excellent
Choosing is made of transparent material, in order to observe the electroelution situation of electroelution cup 2.
Without loss of generality, the present invention can also be provided with voltage-regulation knob 11 on base 1;The work of voltage-regulation knob 11
With being voltage swing for adjusting and controlling to be applied between the electrode slice 4 of the both sides of electroelution cavity 3.Certainly, it is being provided with
During multiple slots 6, can a voltage-regulation knob 11 be set for each slot 6 is corresponding in theory or all of slot 6 is common
With same voltage-regulation knob 11;Situation wherein shown in accompanying drawing 1 shares same voltage-regulation knob for four slots 6
11。
More specifically, corresponding power switch 12 and power supply terminal 13 are typically also provided with base 1;Wherein power supply is opened
The effect for closing 12 is the break-make for controlling the operating circuit of whole electroelution instrument, and power supply terminal 13, then it is by electroelution instrument
The port connected with external power source, thinks that electroelution instrument is powered.Wherein, with the example shown in accompanying drawing 1, its electroelution
Circuit connecting relation inside instrument can refer to and is attached shown in accompanying drawing 10.
Electroelution instrument described in the invention described above, it is typically as follows using process:
1st, the purpose region knife blade after Ago-Gel or polyacrylamide gel electrophoresis is carefully cut, as far as possible
, be then gently transferred to for it in electroelution cavity 3 in electroelution cup 2 by removal redundance;
2nd, electroelution cup 2 is inserted in the slot 6 on electroelution instrument base 1, and ensures the electrode on electroelution cup 2
Electroconductive elastic sheet 7 in piece 4 and slot 6 is in electrical contact good;Then, can be added in electroelution cavity 3 according to subsequent experimental needs
Enter appropriate buffer solution, and gel sample to be eluted should be covered;
3 and then can be closed the lid on corresponding electroelution cup 2, be powered afterwards, and regulation is applied to electroelution cavity
The voltage of 3 both sides carries out wash-out treatment for the constant pressure of 5-10V, and elution time is generally 10min or so;In addition when wash-out target sample
When product are protein, whole electroelution instrument can be also put into 4 DEG C of refrigerator, be carried out at wash-out with being maintained under the temperature environment
Reason;And can before buffer solution is added the first corresponding buffer solution of precooling;
4th, after sample elution is finished, deenergization takes out electroelution cup 2, then carefully suctions out electroelution with pipettor
During buffer solution in cup 2 manages (pipe centrifugation) to EP, purifying or Direct precipitation are then selected with recovery sample according to subsequent experimental.
Electroelution instrument of the present invention at least has following purposes:
1. agarose gel electrophoresis DNA fragmentation is reclaimed.
2. polyacrylamide gel electrophoresis DNA fragmentation is reclaimed.
3. polyacrylamide gel electrophoresis Protein Recovery.
Electroelution instrument of the present invention is contrasted with existing product:
Conventional electroelution device mainly has the D-tube electroelutions pipe and U.S. Bio- of MERCK (Merck) company now
The type electroelution instrument of Bole 422 of Rad (Bole) company.There is much in common in face in both, be all using semi-transparent in design
Film is intercepted to reclaiming fragment, is then that operation can just obtain purpose fragment carrying out one;Wherein, the latter comes with respect to the former
Say, although avoid influence of the buffer solution to subsequent experimental, but need additional extra multiple centrifugal process.The present invention and D-
The performance comparison situation such as following table of tube electroelutions pipe and the type electroelution instrument of Bole 422:
The present invention of table 1 and D-tube electroelutions pipe and the performance comparison of the type electroelution instrument of Bole 422
Claims (7)
1. electroelution instrument, it is characterised in that:Including base (1) and electroelution cup (2);It is internal in the electroelution cup (2)
With electroelution cavity (3);The electrode slice of sheet is respectively arranged with the relative wall of cup in electroelution cavity (3) both sides
, and the one side of the electrode slice (4) exposes from the outer surface of electroelution cup (2) (4);Washed with electricity per plate electrode piece (4)
Connected by the conductive mouth (5) through wall of cup between de- cavity (3);Slot (6) is provided with the base (1), described
The both sides of slot (6) are respectively arranged with electroconductive elastic sheet (7);The electroelution cup (2) is removably inserted into the slot (6)
It is interior, and after the electroelution cup (2) is inserted into the slot (6), the electrode slice (4) can respectively with respective side
Electroconductive elastic sheet (7) electrically conduct.
2. electroelution instrument as claimed in claim 1, it is characterised in that:The profile of the electroelution cup (2) is in cuboid,
And its one end is provided with opening portion (8);The electrode slice (4) is arranged on the one of which phase of electroelution cup (2) surrounding
To wall of cup on.
3. electroelution instrument as claimed in claim 2, it is characterised in that:Electrode slice (4) on wall of cup in specular and
The setting being parallel to each other.
4. electroelution instrument as claimed in claim 2, it is characterised in that:The one end for being provided with opening portion (8) is electroelution cup
(2) top, one end relative with opening portion (8) is the bottom of electroelution cup (2);In the bottom of electroelution cup (2), if
Extension is inclined to middle part after being equipped with the both sides wall of cup bending of electrode slice (4), and V-arrangement ditch is formed in the bottom of electroelution cavity (3)
Slot structure (9) or inverted trapezoidal groove structure (10).
5. electroelution instrument as claimed in claim 2, it is characterised in that:The conductive mouth (5) is elongated, and conductive mouth
(5) length direction is consistent with the depth direction of electroelution cavity (3).
6. electroelution instrument as claimed in claim 1, it is characterised in that:It is provided with base (1) multiple independent of each other slotting
Groove (6).
7. the electroelution instrument as described in any one in claim 1 to 6, it is characterised in that:Electricity is additionally provided with base (1)
Pressure adjusting knob (11).
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CN201710099124.3A CN106754893B (en) | 2017-02-23 | 2017-02-23 | Electrolysis apparatus |
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CN201710099124.3A CN106754893B (en) | 2017-02-23 | 2017-02-23 | Electrolysis apparatus |
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CN106754893B CN106754893B (en) | 2023-07-25 |
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Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4747918A (en) * | 1986-06-18 | 1988-05-31 | William Bellomy | Electroelution apparatus and method of using same |
US4750982A (en) * | 1985-03-21 | 1988-06-14 | Lifecodes Corp. | Process and apparatus for purifying and concentrating DNA from crude mixtures containing DNA |
US6649419B1 (en) * | 2000-11-28 | 2003-11-18 | Large Scale Proteomics Corp. | Method and apparatus for protein manipulation |
CN102051357A (en) * | 2010-11-24 | 2011-05-11 | 同济大学 | Super-micro deoxyribonucleic acid/ribonucleic acid electro-eluter |
CN104807872A (en) * | 2015-04-08 | 2015-07-29 | 边高鹏 | Special electrophoresis tank for comet assay |
CN206751852U (en) * | 2017-02-23 | 2017-12-15 | 四川维度创研生物科技有限公司 | Electroelution instrument |
-
2017
- 2017-02-23 CN CN201710099124.3A patent/CN106754893B/en active Active
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4750982A (en) * | 1985-03-21 | 1988-06-14 | Lifecodes Corp. | Process and apparatus for purifying and concentrating DNA from crude mixtures containing DNA |
US4747918A (en) * | 1986-06-18 | 1988-05-31 | William Bellomy | Electroelution apparatus and method of using same |
US6649419B1 (en) * | 2000-11-28 | 2003-11-18 | Large Scale Proteomics Corp. | Method and apparatus for protein manipulation |
CN102051357A (en) * | 2010-11-24 | 2011-05-11 | 同济大学 | Super-micro deoxyribonucleic acid/ribonucleic acid electro-eluter |
CN104807872A (en) * | 2015-04-08 | 2015-07-29 | 边高鹏 | Special electrophoresis tank for comet assay |
CN206751852U (en) * | 2017-02-23 | 2017-12-15 | 四川维度创研生物科技有限公司 | Electroelution instrument |
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