CN112342125A - Extrusion type biomacromolecule recovery device and recovery method - Google Patents
Extrusion type biomacromolecule recovery device and recovery method Download PDFInfo
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- CN112342125A CN112342125A CN202011252912.XA CN202011252912A CN112342125A CN 112342125 A CN112342125 A CN 112342125A CN 202011252912 A CN202011252912 A CN 202011252912A CN 112342125 A CN112342125 A CN 112342125A
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- 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
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- 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
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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Abstract
The invention discloses an extrusion type biomacromolecule recovery device and a recovery method, wherein the device comprises a recovery pipe, a push rod and a filter membrane; the filter membrane is detachably arranged in the recovery pipe; one end of the push rod is inserted into the recovery pipe in a sliding way and is used for being matched with the filter membrane to extrude an adhesive tape containing biomacromolecules between the push rod and the filter membrane so as to discharge the biomacromolecules at the bottom of the recovery pipe along with the extruded liquid. According to the recovery device and the recovery method, the rubber strips containing the biomacromolecules are extruded through the push rod and the filter membrane, the biomacromolecules with specific molecular weights in the rubber strips can be selectively extruded and recovered according to the pore sizes of the filter membrane, the recovery device does not need to dissolve the rubber strips, the operation is simple, and the recovery time is short.
Description
Technical Field
The invention relates to the technical field of biomacromolecule recovery, in particular to an extrusion type biomacromolecule recovery device and a recovery method.
Background
Biological macromolecules mainly include DNA, RNA, and proteins; however, the recovery of gel cuts from biomacromolecules such as DNA, RNA and proteins is a routine operation in biological laboratories, such as the purification of PCR products, enzyme cuts, etc.
At present, the method for recovering DNA from agarose gel mainly comprises the steps of dissolving agarose gel by guanidine salt, then loading the agarose gel on a centrifugal adsorption column, and then eluting DNA; or putting the gel in an electric field for electroelution; however, the above recovery method requires 30 to 60 minutes for recovery, which is long and cumbersome.
RNA is very easy to degrade, and the complicated operation easily causes the degradation, so that a proper recovery method is not available.
The medium for protein electrophoresis is mainly polyacrylamide gel, which cannot be dissolved because of covalent connection; the existing recovery method is mainly electroelution, however, the electroelution has the problem of longer time.
Disclosure of Invention
The invention aims to provide an extrusion type biomacromolecule recovery device and a recovery method.
In order to achieve the above purpose of the present invention, the following technical solutions are adopted:
the invention provides an extrusion type biomacromolecule recovery device, which comprises a recovery pipe, a push rod and a filter membrane;
the filter membrane is detachably arranged in the recovery pipe;
one end of the push rod is inserted into the recovery pipe in a sliding way and is used for being matched with the filter membrane to extrude an adhesive tape containing biomacromolecules between the push rod and the filter membrane so as to discharge the biomacromolecules at the bottom of the recovery pipe along with the extruded liquid.
The recovery device provided by the invention extrudes the adhesive tape containing the biomacromolecules through the push rod and the filter membrane, can selectively extrude and recover the biomacromolecules with specific molecular weights in the adhesive tape according to the pore size of the filter membrane, does not need to dissolve the adhesive tape, and is simple to operate and short in recovery time.
Preferably, the end of the push rod is provided with a squeezing pad, and the squeezing pad can be tightly attached to the inner wall of the recovery pipe.
Through the setting of extrusion pad can make push rod low side and recovery tube inner wall sealing connection, extrusion adhesive tape that can be better, more abundant extrude the biomacromolecule in the adhesive tape, improve the rate of recovery.
Preferably, the interior of the recovery tube is sequentially divided into an extrusion area, a transition area and a leakage opening from top to bottom, the extrusion area and the leakage opening are cylindrical, the diameter of the extrusion area is larger than that of the leakage opening, the transition area is conical, and the filter membrane is located at the bottom of the extrusion area.
Preferably, the device further comprises a gasket, the gasket is positioned at the bottom of the filter membrane, and a plurality of through holes are formed in the gasket.
Through the setting of gasket, prevent that extrusion in-process filter membrane warp, lead to the extrusion not thorough, and then can improve the rate of recovery of biomacromolecule, in addition, can also prevent and treat that the filter membrane is damaged in extrusion process.
Preferably, the biomacromolecule is at least one of DNA, RNA, protein and lipopolysaccharide.
The second aspect of the invention provides a biomacromolecule recovery method based on the device, wherein the recovery method comprises the following steps:
placing the filter membrane in a recovery tube, and then placing the rubber strip containing the biomacromolecules in the recovery tube;
and inserting the push rod into the recovery pipe and extruding the rubber strip containing the biomacromolecules to discharge the biomacromolecules at the bottom of the recovery pipe.
The recovery method is simple to operate, can quickly recover the biomacromolecules in the adhesive tape, and has excellent recovery rate.
Preferably, the biomacromolecule is at least one of DNA, RNA and protein.
Preferably, the gasket is placed in the recovery tube before the filter membrane is placed in the recovery tube.
Compared with the prior art, the invention has the beneficial effects that at least:
the recovery device provided by the invention extrudes the adhesive tape containing the biomacromolecules through the push rod and the filter membrane, can selectively extrude and recover the biomacromolecules with specific molecular weights in the adhesive tape according to the pore size of the filter membrane, does not need to dissolve the adhesive tape, and is simple to operate and short in recovery time.
Drawings
In order to more clearly illustrate the detailed description of the invention or the technical solutions in the prior art, the drawings that are needed in the detailed description of the invention or the prior art will be briefly described below. Throughout the drawings, like elements or portions are generally identified by like reference numerals. In the drawings, elements or portions are not necessarily drawn to scale.
FIG. 1 is a schematic view of an extrusion-type bio-macromolecule recycling device provided by an embodiment of the present invention;
FIG. 2 shows the results of gel running before and after extrusion recovery of biomacromolecule DNA in the experimental example of the present invention;
FIG. 3 shows the results of gel running before and after extrusion recovery of biomacromolecule RNA in the experimental example of the present invention;
FIG. 4 shows the results of gel running before and after extrusion recovery of biopolymer proteins in the experimental example of the present invention;
FIG. 5 shows the results of gel running before and after extrusion recovery of biopolymer lipopolysaccharide in the experimental example of the present invention.
Shown in the figure:
1-recovery tube, 11-extrusion zone, 12-transition zone, 13-leakage orifice; 2-push rod, 21-extrusion pad; 3-a filter membrane; 4-adhesive tape.
Detailed Description
The following describes embodiments of the present invention in detail with reference to the following embodiments. The following examples are only for illustrating the technical solutions of the present invention more clearly, and therefore are only examples, and the protection scope of the present invention is not limited thereby.
It is to be noted that, unless otherwise specified, technical or scientific terms used herein shall have the ordinary meaning as understood by those skilled in the art to which the invention pertains.
The embodiment of the invention provides an extrusion type biomacromolecule recovery device, which comprises a recovery pipe 1, a push rod 2 and a filter membrane 3, wherein the push rod is connected with the recovery pipe 1;
the filter membrane 3 is detachably arranged in the recovery pipe 1;
one end of the push rod 2 is slidably inserted into the recovery tube 1 and is used for extruding a rubber strip 4 containing biomacromolecules between the push rod and the filter membrane 3 in a matching way so as to discharge the biomacromolecules at the bottom of the recovery tube 1 along with the extruded liquid.
According to the recovery device, the rubber strips 4 containing biological macromolecules are extruded through the push rod 2 and the filter membrane 3, the biological macromolecules with specific molecular weights in the rubber strips 4 can be selectively extruded and recovered according to the pore size of the filter membrane 3, the recovery device does not need to dissolve the rubber strips 4, the operation is simple, and the recovery time is short.
In one embodiment, the end of the push rod 2 is provided with a pressing pad 21, and the pressing pad 21 can be closely attached to the inner wall of the recovery pipe 1.
Can make 2 low ends of push rod and 1 inner wall sealing connection of recovery tube through the setting of extrusion pad 21, extrusion adhesive tape 4 that can be better, more abundant extrude the biomacromolecule in adhesive tape 4, improve the rate of recovery of biomacromolecule.
The specific structure of the recovery tube 1 is not strictly limited in the present invention, for example, in one embodiment, the interior of the recovery tube 1 is divided into an extrusion region 11, a transition region 12 and a leakage opening 13 from top to bottom in sequence, the extrusion region 11 and the leakage opening 13 are cylindrical, the diameter of the extrusion region 11 is larger than that of the leakage opening 13, the transition region 12 is conical, and the filter membrane 3 is located at the bottom of the extrusion region 11.
In one embodiment, the above recycling device of the present invention further comprises a gasket (not shown), the gasket is located at the bottom of the filter membrane 3, and the gasket is provided with a plurality of through holes.
Through the setting of gasket, 3 supporting role of filter membrane can be given to the gasket, prevent to extrude in-process filter membrane 3 deformation, lead to the extrusion not thorough, and then can improve the rate of recovery of biomacromolecule, in addition, can also prevent and treat that filter membrane 3 is damaged in extrusion process.
The present invention is not strictly limited to biological macromolecules, for example, biological macromolecules may be at least one of DNA, lipopolysaccharide, RNA, and protein; in one embodiment, the biological macromolecule may be DNA; in other embodiments, the biomacromolecule may be a mixture of DNA and protein.
The embodiment of the invention also provides a biomacromolecule recovery method based on the device, which comprises the following steps:
placing the filter membrane in a recovery tube, and then placing the rubber strip containing the biomacromolecules in the recovery tube;
and inserting the push rod into the recovery pipe and extruding the rubber strip containing the biomacromolecules to discharge the biomacromolecules at the bottom of the recovery pipe.
The recovery method is simple to operate, can quickly recover the biomacromolecules in the adhesive tape, and has excellent biomacromolecule recovery rate.
In one embodiment, the spacer is placed in the recovery tube prior to placing the filter membrane in the recovery tube; the filter membrane is supported by adding a spacer.
According to the recovery device and the recovery method, the rubber strips containing the biomacromolecules are extruded through the push rod and the filter membrane, the biomacromolecules with specific molecular weights in the rubber strips can be selectively extruded and recovered according to the pore sizes of the filter membrane, the recovery device does not need to dissolve the rubber strips, the operation is simple, and the recovery time of the biomacromolecules is short.
Experimental example 1
Experimental materials:
the Marker DNA Ladder is formed by mixing separately prepared PCR products, has 6 DNA fragments in total, is added with a sample buffer solution, and can be directly electrophoresed; for the convenience of post-electrophoresis observation, the specificity was enhanced by a 400bp band at a concentration of about 100 ng/5. mu.l, and the DNA concentration of the other bands was about 50 ng/5. mu.l.
lambda DNA (0.35ug/uL 21023 cargo space)
Human genomic DNA
RNA
Experimental methods and results:
performing electrophoresis gel running on the Marker, the human genome DNA and the lambda DNA respectively, wherein the electrophoresis conditions are shown in table 1, and the gel running results are shown in the left picture in fig. 2, wherein the human genome DNA and the lambda DNA are cut off from the gel strip after electrophoresis, the extrusion type biomacromolecule recovery device shown in fig. 1 is adopted for extrusion, an extrusion solution containing the human genome DNA and the lambda DNA is recovered, then, the Marker, the extrusion solution containing the human genome DNA and the extrusion solution containing the lambda DNA are subjected to electrophoresis gel running, and the gel running results are shown in the right picture in fig. 2;
as can be seen from FIG. 2, the recovery method based on the extrusion-type biomacromolecule recovery apparatus of the present invention enables recovery of biomacromolecules (DNA) in the rubber strip.
Performing electrophoresis gel running on a Marker and RNA respectively, wherein the electrophoresis conditions are shown in table 1, and the gel running result is shown in the left picture in figure 3, wherein the gel strip containing RNA is cut off from the gel strip after electrophoresis, the extrusion type biomacromolecule recovery device shown in figure 1 is adopted for extrusion, the extrusion liquid containing RNA is recovered, then, the Marker and the extrusion liquid containing RNA are subjected to electrophoresis gel running, and the gel running result is shown in the right picture in figure 3;
as can be seen from FIG. 3, the recovery method based on the extrusion-type biopolymer recovery device of the present invention enables recovery of the biopolymer (RNA) in the adhesive tape.
TABLE 1 conditions of electrophoresis
Experimental example 2
The experimental example is a protein prestained Marker (25kDa)
Run 8. mu.l protein Prestain marker (10-180kDa) first, electrophoresis conditions: the voltage is 90V, the current is 15mA, and the time is 1 hour and 20 minutes; the electrophoresis results are shown in the left panel of FIG. 4;
cutting a 25kDa strip by using a blade after the glue running is finished; then, the extrusion type biomacromolecule recovery device shown in the figure 1 is adopted to carry out extrusion recovery to obtain liquid;
adding a proper amount of loading buffer solution into the obtained liquid, dropping the whole liquid into a glue hole, and simultaneously dropping 8 mu l of marker (10-180kDa) for electrophoresis, wherein the electrophoresis conditions are as follows: voltage: the voltage is 90V, the current is 15mA, and the time is 1 hour and 20 minutes; the electrophoresis results are shown in the right panel of FIG. 4;
as can be seen from fig. 4:
the recovery method based on the extrusion type biomacromolecule recovery device can recover biomacromolecules (proteins) in the adhesive tape.
Experimental example 3
Run 5. mu.l protein marker (10-180kDa) and 2. mu.l lipopolysaccharide solution (5mg/ml), voltage: 70V, 15mA of current, 1 hour and 40 minutes.
After running the gel, the gel was stained and the results are shown in the left panel of fig. 5;
cutting a band of lipopolysaccharide with a blade; the liquid is obtained by extrusion and recovery by using the extrusion type biomacromolecule recovery device shown in figure 1.
To the resulting liquid was added an appropriate amount of loading buffer, and the whole liquid was spotted into the wells while 5. mu.l of marker, voltage: 70V, 15mA of current, 1 hour and 40 minutes; after running the gel, the gel was stained and the results are shown in the right panel of FIG. 5;
as can be seen from fig. 5:
the recovery method based on the extrusion type biomacromolecule recovery device can recover biomacromolecules (lipopolysaccharides) in the adhesive tape.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; such modifications and substitutions do not depart from the spirit and scope of the present invention, and they should be construed as being included in the following claims and description.
Claims (8)
1. An extrusion type biomacromolecule recovery device is characterized by comprising a recovery pipe, a push rod and a filter membrane;
the filter membrane is detachably arranged in the recovery pipe;
one end of the push rod is inserted into the recovery pipe in a sliding way and is used for being matched with the filter membrane to extrude an adhesive tape containing biomacromolecules between the push rod and the filter membrane so as to discharge the biomacromolecules at the bottom of the recovery pipe along with the extruded liquid.
2. The device of claim 1, wherein the end of the push rod is provided with a squeezing pad capable of closely fitting with the inner wall of the recovery pipe.
3. The device as claimed in claim 1, wherein the interior of the recovery tube is divided into an extrusion area, a transition area and a leakage opening from top to bottom in sequence, the extrusion area and the leakage opening are cylindrical, the diameter of the extrusion area is larger than that of the leakage opening, the transition area is conical, and the filter membrane is positioned at the bottom of the extrusion area.
4. The device of claim 1, further comprising a spacer positioned at the bottom of the filter membrane, the spacer having a plurality of through holes disposed therein.
5. The device of claim 1, wherein the biomacromolecule is at least one of DNA, RNA, lipopolysaccharide, and protein.
6. A method for recovering biological macromolecules based on the device of any one of claims 1 to 5, comprising the steps of:
placing the filter membrane in a recovery tube, and then placing the rubber strip containing the biomacromolecules in the recovery tube;
and inserting the push rod into the recovery pipe and extruding the rubber strip containing the biomacromolecules to discharge the biomacromolecules at the bottom of the recovery pipe.
7. The recovery method according to claim 6, wherein the biomacromolecule is at least one of DNA, RNA, protein and lipopolysaccharide.
8. The method of claim 6, wherein the spacer is placed in the recovery tube before the filter membrane is placed in the recovery tube.
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| CN202011252912.XA CN112342125A (en) | 2020-11-11 | 2020-11-11 | Extrusion type biomacromolecule recovery device and recovery method |
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| CN202011252912.XA CN112342125A (en) | 2020-11-11 | 2020-11-11 | Extrusion type biomacromolecule recovery device and recovery method |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020040872A1 (en) * | 2000-07-18 | 2002-04-11 | Bogoev Roumen A. | Device and methods for subdividing and filtering gel material and extracting molecules therefrom |
| CN103284885A (en) * | 2013-06-28 | 2013-09-11 | 陈庆强 | Medicine feeding injector with filtering device |
| CN107441771A (en) * | 2017-08-18 | 2017-12-08 | 杭州立昂科技有限公司 | The interaction of divided porous particulate and liquid and separated in filtering outlet syringe |
| CN211079163U (en) * | 2019-08-05 | 2020-07-24 | 湖州河马生物科技有限公司 | Anti-blocking probe for DNA sampling |
-
2020
- 2020-11-11 CN CN202011252912.XA patent/CN112342125A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020040872A1 (en) * | 2000-07-18 | 2002-04-11 | Bogoev Roumen A. | Device and methods for subdividing and filtering gel material and extracting molecules therefrom |
| CN103284885A (en) * | 2013-06-28 | 2013-09-11 | 陈庆强 | Medicine feeding injector with filtering device |
| CN107441771A (en) * | 2017-08-18 | 2017-12-08 | 杭州立昂科技有限公司 | The interaction of divided porous particulate and liquid and separated in filtering outlet syringe |
| CN211079163U (en) * | 2019-08-05 | 2020-07-24 | 湖州河马生物科技有限公司 | Anti-blocking probe for DNA sampling |
Non-Patent Citations (1)
| Title |
|---|
| 薛仁镐等: "琼脂糖凝胶中DNA 片段的挤压回收法", 《生物技术》 * |
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Application publication date: 20210209 |