CN102586225A - Method for separating target molecules by controlling magnetic beads - Google Patents
Method for separating target molecules by controlling magnetic beads Download PDFInfo
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- CN102586225A CN102586225A CN2011100078820A CN201110007882A CN102586225A CN 102586225 A CN102586225 A CN 102586225A CN 2011100078820 A CN2011100078820 A CN 2011100078820A CN 201110007882 A CN201110007882 A CN 201110007882A CN 102586225 A CN102586225 A CN 102586225A
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Abstract
The invention discloses a method for separating target molecules by controlling magnetic beads. The method comprises the following steps: (1) binding: placing the magnetic beads, liquor containing the target molecules, and binding buffer liquor into a container and mixing, so that the magnetic beads and the target molecules are bound to form a composite; and removing impurities and remaining the composite in the container; (2) washing: adding washing buffer liquor into the container so as to mix the washing buffer liquor and the composite, removing impurities and remaining the composite in the container; and (3) eluting: adding the washing buffer liquor into the container so as to mix the washing buffer liquor with the composite, separating the container from a magnetic field, incubating to separate the magnetic beads from the target molecules and collecting the target molecules. The test proves that the yield of the method is higher than that of the traditional suction method and the consistency of centrifugal tubes is more excellent than that in the traditional suction method. Therefore, the method has a wide application prospect in the fields of separation and purification of biological molecules.
Description
Technical field
The present invention relates to a kind of method of handling magnetic bead separate targets molecule.
Background technology
A large amount of Bioexperiment and clinical diagnosis be unable to do without specimen preparation, and the extraction of biomolecules and purification technique extensive application in specimen preparation.Along with development of technologies, the handled sample size of clinical diagnosis and Bioexperiment is also increasing.
It is specimen preparation a kind of technology commonly used that magnetic bead is handled.Present magnetic bead mixing and stripping technique mainly contain two types: one type is to collect, discharge, shift through the relative movement realization magnetic bead of bar magnet and magnetosheath, and then the extraction of completion biomolecules.The physical construction relative complex that this method adopted exist to realize the high problem of robotization cost, and magnetosheath is as consumptive material, and price is higher.Simultaneously, because magnetosheath fully contacts with sample, there is the risk of contaminated samples, and in the magnetic bead transfer process, has the problem of magnetic bead loss.In addition, because the motion of magnetosheath is comparatively violent, the integrity of the artifact molecule that can't guarantee to purify.
Other one type is rifle head suction mode, promptly uses rifle head pressure-vaccum mixing, and magnet absorption magnetic bead separates to do magnetic bead.This method realizes that the cost of robotization is high, needs several robotic arms and pump, complex structure.In addition, also exist disposable processing sample limited amount in the problem of mechanical arm.In a word, need raise the efficiency and many factors such as disposable processing sample size, with satisfy low consumptive material, high efficiency biomolecules is extracted and the purifying requirement.
Summary of the invention
The purpose of this invention is to provide a kind of manipulation magnetic bead with target molecule method of separating from solution, specifically is a kind ofly to make combination, washing and separate more fully, makes the target molecule purity and the higher magnetic bead separation method of yield that obtain.
The magnetic bead that utilizes of the present invention comprises the steps: target molecule isolating method from solution
(1) combines: place container to mix magnetic bead, the solution that contains target molecule and binding buffer liquid; With said container place the magnetic force scope in magnetic field and make said container magnetic bead magnetic field force induced direction and/or size be in continuous change state; So that the fully contact each other of all materials in the said container, magnetic bead combines to form mixture with target molecule; Remove impurity, keep mixture in said container;
(2) washing: in said container, add lavation buffer solution; Lavation buffer solution is mixed with said mixture; With said container place the magnetic force scope in magnetic field and make said container magnetic bead magnetic field force induced direction and/or size be in continuous change state so that each other fully contact of all materials in the said container; Remove impurity, keep mixture in said container;
(3) wash-out: in said container, add elution buffer; Elution buffer is mixed with said mixture; With said container place the magnetic force scope in magnetic field and make said container magnetic bead magnetic field force induced direction and/or size be in continuous change state so that each other fully contact of all materials in the said container; Again said container is broken away from magnetic field, hatch,, collect target molecule so that said magnetic bead separates with said target molecule.
In the said process, in said step (1), (2) or (3), said make magnetic bead magnetic field force induced direction and/or the size method that is in continuous change state specifically can be shown in following I, II or the III, but do not get rid of other method;
I, make said container and the relative movement of said magnetic field, the magnet that wherein produces magnetic field is permanent magnet;
The size and/or the direction of II, change magnetic field force, the magnet that wherein produces magnetic field is electro-magnet; Specifically can the magnetic force size be changed through changing the size of current of electromagnet; Change the current of electromagnet direction, make S the utmost point change.
III, change the size and/or the direction of magnetic field force and make said container and the relative movement of said magnetic field, the magnet that wherein produces magnetic field is electro-magnet.
In the said process, the said method of said container and the relative movement of said magnetic field that makes specifically can be following 1), 2) or 3) shown in:
1) said container is taken exercises between two opposite rows magnet;
2) said container is around said magnet movement;
3) said magnet moves around said container.
In the said process, said 1), 2) or 3) in, said motion can be circumferential motion, uniform motion or uniform circular motion.
In the said process, the speed of said uniform circular motion is 8rpm-75rpm, is preferably 12rpm-30rpm or 15rpm-30rpm, is specially 15rpm, 19rpm, 23rpm or 30rpm.
If movement velocity can let magnetic bead also have little time motion in pipe too soon, got back to original position again, not only can not be allowed to condition at sufficient movement in the pipe, may destroy the integrity of original biomolecules in addition; If movement velocity is too slow, efficient again can be very low and can not lets magnetic bead fully scatter to touch liquid.
In the said process, during said container is taken exercises between two opposite rows magnet, the said container comb that several independent pipe coupling form of serving as reasons, and said comb is straight line.
In the said process,, keep preferably that each independent pipe causes at the magnetic field force induced homogeneous of synchronization in the said comb in order to make the effect in each pipe consistent.
In the said process; Make each independent pipe in the said comb synchronization the method that causes of magnetic field force induced homogeneous specifically can be as follows; But be not limited thereto method: every row's magnet is connected in sequence by plurality of cuboid shape permanent magnet; Permanent magnet closure linearly and perpendicular to the N utmost point and the S utmost point line direction of permanent magnet self, the pole orientation of two adjacent permanent magnets is opposite; Two relative between two row's magnet permanent magnet edge lines are rectangular or square, and the magnetic pole of two relative permanent magnets is identical; Comb and two row's magnet equal keeping parallelisms, each rectangular-shaped permanent magnet equals the width of each single tube along the widest part of the tangent plane of the N utmost point of magnet and S utmost point line direction along the width of the tangent plane of himself N utmost point and S utmost point line direction; And comb is at the non-fringing effect regional movement of two row's magnet, and said fringing effect is the fringing effect that the two ends of every row's magnet produce.
In the said process, said independent pipe is test tube or centrifuge tube; The said comb that is formed by several independent pipe coupling is eight platoon centrifuge tubes.
In the said process, said target molecule is a biomolecules; Or said biomolecules is DNA, RNA, glucide or protein.
The inventive method through change magnetic bead magnetic field force induced size and Orientation, magnetic bead combined with target molecule more fully or separate, finally obtain target molecule.Specifically, can take respectively following four kinds of modes change magnetic bead magnetic field force induced size and Orientation: 1) magnet is fixed, and the motion of magnetic bead system; 2) the magnetic bead system is fixed, and magnet movement; 3) magnetic bead system and magnet all move; 4) adopt electro-magnet conversion magnetic force size and Orientation.
Said magnet can be electro-magnet, also can be permanent magnet.
Said permanent magnet mainly is to be used to provide magnetic field, and through regulate and sample between distance and the motion of direction with the control magnetic bead, thereby the suffered magnetic field force size and Orientation of change magnetic bead system.
Said electro-magnet mainly is through regulating self size of current and direction, and combine to regulate and sample between distance and direction, thereby change the suffered magnetic field force size and Orientation of magnetic bead system.
In the inventive method, the effect that the magnetic field force induced size and Orientation of magnetic bead is changed is specific as follows:
1, combines: make container that the magnetic bead system is housed be in continuous change state with respect to the distance and/or the position of magnet; Make magnetic bead under the effect of the magnetic field force that changes in system sufficient movement; And then make the abundant mixing of magnetic bead and biomolecules and contact, form the mixture of magnetic bead and biomolecules.
2, washing: container is pressed close to magnet for some time; After removing unnecessary liquid; Washings is added in the magnetic bead be combined, makes container that the magnetic bead system is housed be in continuous change state with respect to the distance and/or the position of magnet, make magnetic bead under the effect of the magnetic field force that changes in system sufficient movement; And then make the abundant mixing of mixture and washings and fully contact, thereby make it obtain thorough washing.
3, wash-out: container is pressed close to magnet for some time, remove unnecessary liquid after, elutriant is added in the good magnetic bead of washing; Make container that the magnetic bead system is housed be in continuous change state with respect to the distance and/or the position of magnet; The control magnetic bead moves in elutriant, make magnetic bead under the effect of the magnetic field force that changes in system sufficient movement, make the abundant mixing of mixture and elutriant and contact; Thereby be beneficial to wash-out, thereby obtain high density, highly purified target molecule.
The present invention is owing to take above technical scheme, and it has the following advantages:
1) because centrifuge tube adopts slower movement velocity, the suffered shearing force of target biological molecules is less, and is difficult for producing bubble, the biomolecules integrity after guaranteeing to purify.
2) absence of liq is spattered phenomenon outward in the manipulation process, has reduced the crossed contamination possibility.
3) blending process does not have other experiment consumptive material intervention, when reducing cost, has reduced possible pollution.
4) can improve the flux of biomolecules extraction and purifying with a plurality of samples as for carrying out mixing in the magnetic field.
5) only need the relative position of regulating magnet and magnetic bead system, or the magnetic force of conversion electro-magnet size, it is simple relatively to move, thus easily be automated, and cost is low.
6) do not need special container, only need common consumptive material, practicality is very strong
7) whole process is not carried out the magnetic bead transfer, has reduced the magnetic bead loss.
The biomolecules extraction yield that experiment showed, the inventive method is higher than traditional suction mode, and the consistence between multitube is better than tradition suction mode.Therefore, the inventive method will have broad application prospects in the specimen preparation field of biomolecules.
Description of drawings
Fig. 1 is the synoptic diagram of the embodiment of the invention one.
Fig. 2 is the synoptic diagram of the embodiment of the invention one.
Fig. 3 is the synoptic diagram of the embodiment of the invention two.
Fig. 4 is the synoptic diagram of the embodiment of the invention two.
Fig. 5 is the synoptic diagram of the embodiment of the invention three.
Fig. 6 is the synoptic diagram of the embodiment of the invention three.
Fig. 7 is the synoptic diagram of the embodiment of the invention three.
Embodiment
Employed experimental technique is ordinary method like no specified otherwise among the following embodiment.
Used material, reagent etc. like no specified otherwise, all can obtain from commercial sources among the following embodiment.
In the accompanying drawing, each mark is following: 1, magnetic bead system; 2, magnet.
Damping fluid GD among the following embodiment, rinsing liquid PW, elutriant TB, magnetic bead suspension B are product in the paramagnetic particle method genome DNA extracting reagent kit.The paramagnetic particle method genome DNA extracting reagent kit is available from TIANGEN Biotech (Beijing) Co., Ltd., and catalog number is DP329.
The kind of the solution that each step need add in the experiment and consumption are operated according to the specification sheets of paramagnetic particle method genome DNA extracting reagent kit.
The PCR product that following embodiment 1,2,3,4 uses is identical same a collection of PCR product, and what embodiment 5 used is another batch PCR product.All contain in the PCR product with magnetic bead bonded DNA (promptly treating separated DNA).
A centrifuge tube and a magnet are one group, and magnet is fixed, and centrifuge tube is done uniform circular motion round magnet.Magnet is the monolithic permanent magnet, and the monolithic permanent magnet is rectangular-shaped (Fig. 1 and 2).Centrifuge tube all is in moving process in the magnetic force scope in the magnetic field that magnet produces.
One, combines
1, preparation system:
1) the PCR product of adding 40 μ L in each pipe of 3 groups of centrifuge tubes;
2) in each pipe, add 10 μ L damping fluid GD again;
3) mixed solution of adding 110 μ L PW and 20 μ L absolute ethyl alcohols in each pipe, the vortex mixing;
4) add 5 μ L, 10 μ L or 15 μ L magnetic bead suspension B respectively in three groups of centrifuge tubes;
2, binding operation
Single centrifuge tube is done uniform circular motion (like Fig. 1, shown in 2) with the speed of 15rpm around permanent magnet; Magnetic bead fully contacts with target molecule and forms mixture in this process; Move after 2 to 3 minutes, stop, stop that mixture is adsorbed onto on the tube wall of container after 30 seconds by magnet near magnet; With liquid-transfering gun liquid is removed, kept mixture;
Two, washing
1, washing for the first time
In the centrifuge tube that mixture is housed that step 1 obtains, add 200 μ L damping fluid GD; Centrifuge tube is done uniform circular motion with the speed of 15rpm around permanent magnet, and magnetic bead fully contacts with damping fluid GD in this process, moves after 2 to 3 minutes; Stop near magnet; Stop that mixture is adsorbed onto on the tube wall by magnet after 30 seconds, liquid is removed, keep mixture with liquid-transfering gun;
2, washing for the second time
In the centrifuge tube of step 1, add 200 μ L rinsing liquid PW; Centrifuge tube is done uniform circular motion with the speed of 15rpm around permanent magnet; Move after 2 to 3 minutes, stop, stop that mixture is adsorbed onto on the tube wall of consumptive material after 30 seconds by magnet near magnet; With liquid-transfering gun liquid is removed, kept mixture; Room temperature was dried 5 minutes.
Three, wash-out
The elutriant TB that in the centrifuge tube of step 2, adds 50～100 μ L; Centrifuge tube is done uniform circular motion with the speed of 15rpm around permanent magnet; Move after 2 to 3 minutes, centrifuge tube is removed magnetic field (promptly leaving the magnetic force scope in magnetic field), place 56 ℃ to hatch 10 minutes centrifuge tube again.Centrifuge tube is stopped near magnet again, stop after 30 seconds, magnetic bead is adsorbed onto on the tube wall by magnet, and target molecule exists and (be target molecule solution) in the dissociating buffer, and target molecule solution is transferred to new pipe.
Four, the detection method of yield
Method: carry out the nucleic acid concentration test with spectrophotometer, attention needs with the Tip head the nucleic acid mixing before the test.
Method and embodiment 1 are basic identical, and different is in combination, washing and elution step, and centrifuge tube is different with the relative movement mode of permanent magnet, is specially: permanent magnet is done uniform circular motion (like Fig. 3, shown in 4) with the speed of 15rpm around centrifuge tube.Be specially: a centrifuge tube and a magnet are one group, and centrifuge tube is fixed, and magnet is done uniform circular motion round centrifuge tube.Magnet is the monolithic permanent magnet, and the monolithic permanent magnet is rectangular-shaped (Fig. 3 and 4).Magnet need guarantee that in moving process centrifuge tube is in the scope of the magnetic force that magnet produces.
Embodiment 3, utilize magnetic bead that target molecule isolating method-container from solution is moved between the magnet two rows
Method and embodiment 1 are basic identical, and different is in combination, washing and elution step, and centrifuge tube is different, specific as follows with the relative movement mode of permanent magnet:
The permanent magnet that two rows are relative and parallel and row's centrifuge tube are one group.One row's centrifuge tube is 8 independent centrifuge tubes that straight line is formed by connecting.The relative magnet positions of two rows is fixed, and eight centrifuge tubes are done as a whole row between the magnet two and done uniform circular motion (Fig. 5,6,7) with the speed of 15rpm.Uniform circular motion can pass through existing techniques in realizing; As eight centrifuge tubes are placed a corresponding rotor; Rotor carries out uniform circular motion by electric motor driving; Thereby eight centrifuge tubes are also done uniform circular motion, and in whole motion process, the relative position of eight centrifuge tubes and rotor is constant.One row's centrifuge tube all is in moving process in the magnetic force scope of two row's magnet generations.
Magnet is permanent magnet, and monolithic magnet is rectangular-shaped; Every row's magnet is to be close to successively to arrange by onesize monolithic permanent magnet to form.
In order to guarantee in eight centrifuge tubes that each pipe at the stressed consistence of synchronization (i.e. each pipe magnetic field force induced size and Orientation unanimity), is provided with magnet arrangement and eight centrifuge tubes are moved according to following mode according to following mode:
Monolithic magnet is close to successively is arranged in a row, monolithic magnet array direction linearly and perpendicular to the N utmost point and the S utmost point line direction of monolithic magnet self, the pole orientation of two adjacent monolithic magnet is opposite; Two relative between two row's magnet monolithic magnet edges lines are rectangular, and the magnetic pole of two relative monolithic magnet is identical; No matter place in the moment or the moving process, all keep comb and two row's magnet opposing parallel at comb in the initial of comb; Each rectangular-shaped permanent magnet equals the width of each single tube along the widest part of the tangent plane of the N utmost point of magnet and S utmost point line direction along the width of the tangent plane of himself N utmost point and S utmost point line direction; Also will make comb the non-fringing effect regional movement of two rows between magnet (fringing effect that produces with the two ends of avoiding two row's magnet makes the unbalance stress of pipe), said fringing effect is meant the fringing effect that the two ends of every row's magnet produce.
The two row's magnet that so are provided with are in non-fringing effect zone; The magneticstrength incomplete same though (magneticstrength of the connecting place of general two blocks of magnet is better than portion other than connected portion of magnet) of each point on the direction parallel with magnet; But because the widest part of the profile of each pipe equates with the width of the profile of magnet; So no matter which comb moves to, its stressing conditions at each pipe of synchronization is identical.
Avoid above-mentioned fringing effect can pass through existing techniques in realizing, specifically can be like the number of the monolithic magnet number of every row's magnet each single tube in the comb, and make the region intermediate motion of comb at every row's magnet, and keep off the two ends of every row's magnet.
So be provided with, just guaranteed that each pipe is all identical at the suffered magnetic field force size and Orientation of synchronization in eight centrifuge tubes, thereby guaranteed consistence between pipe.
In addition, experiment shows, when each monolithic magnet is apart from one another by a segment distance in every row's magnet; The dispersity of the magnetic bead in each centrifuge tube is obviously different; Explain that each pipe is different in the suffered magneticstrength of synchronization in such cases, therefore, consistence places not as above-mentioned being close between pipe.
Experiment also shows; When the pole orientation of two adjacent in every row's magnet monolithic magnet is identical; The dispersity of the magnetic bead in each centrifuge tube is obviously different; Explain that each pipe is different in the suffered magneticstrength of synchronization in such cases, therefore, consistence places not as above-mentioned adjacent magnets pole orientation on the contrary between pipe.
All established 3 groups of experiments in the foregoing description, can the same reaction of N (N >=1) group have been arranged according to demand in the practical application, raised the efficiency.
Embodiment 4, contrast (traditional way)
1, the PCR product (identical with PCR product described in the embodiment 1-3) that in each pipe of 3 groups of centrifuge tube combs, adds 40 μ L for criticizing together
2, in each pipe, add 10 μ L damping fluid GD again;
3, the mixed solution that in each pipe, adds 110 μ L PW and 20 μ L absolute ethyl alcohols, the vortex mixing;
4, add 5 μ L, 10 μ L, 15 μ L magnetic bead suspension B respectively in three groups of centrifuge tubes;
5, centrifuge tube is positioned over left standstill on the magnetic force frame 30 seconds, the careful liquid of removing when treating that magnetic bead adsorbs fully;
6, centrifuge tube is taken off from the magnetic force frame, added 200 μ L rinsing liquid GD, the vortex mixing.
7, centrifuge tube is positioned over left standstill on the magnetic force frame 30 seconds, the careful liquid of removing when treating that magnetic bead adsorbs fully.
8, add 200 μ L rinsing liquid PW, vortex mixing.
9, centrifuge tube is positioned over left standstill on the magnetic force frame 30 seconds, the careful liquid of removing when treating that magnetic bead adsorbs fully, room temperature was dried 5 minutes.
10, add the elutriant TB of 50～100 μ L,, hatched 10 minutes for 56 ℃ according to the mode mixing of step 5.
11, centrifuge tube was pressed close to magnet 30 seconds, treat carefully dna solution to be transferred to new pipe when magnetic bead adsorbs fully, and measure.
The result is as shown in table 1.
Table 1, product reclaim the result
From table, can find out how many amounts that no matter adds magnetic bead is, the yield of the inventive method all is higher than the yield of traditional suction method, and consistence is better than tradition suction mode between pipe.
Embodiment 5, different motion speed
Method: with identical described in the embodiment 3; The speed of different is uniform circular motion is 23rpm and 30rpm.
Result: as shown in table 2.
Table 2, product reclaim the result
Claims (10)
1. one kind is utilized magnetic bead with target molecule isolating method from solution, comprises the steps:
(1) combines: place container to mix magnetic bead, the solution that contains target molecule and binding buffer liquid; With said container place the magnetic force scope in magnetic field and make said container magnetic bead magnetic field force induced direction and/or size be in continuous change state; So that the fully contact each other of all materials in the said container, magnetic bead combines to form mixture with target molecule; Remove impurity, keep mixture in said container;
(2) washing: in said container, add lavation buffer solution; Lavation buffer solution is mixed with said mixture; With said container place the magnetic force scope in magnetic field and make said container magnetic bead magnetic field force induced direction and/or size be in continuous change state so that each other fully contact of all materials in the said container; Remove impurity, keep mixture in said container;
(3) wash-out: in said container, add elution buffer; Elution buffer is mixed with said mixture; With said container place the magnetic force scope in magnetic field and make said container magnetic bead magnetic field force induced direction and/or size be in continuous change state so that each other fully contact of all materials in the said container; Again said container is broken away from magnetic field, hatch,, collect target molecule so that said magnetic bead separates with said target molecule.
2. method according to claim 1 is characterized in that: in said step (1), (2) or (3), said make magnetic bead magnetic field force induced direction and/or the size method that is in continuous change state be shown in following I, II or the III:
I, make said container and the relative movement of said magnetic field, the magnet that wherein produces magnetic field is permanent magnet;
The size and/or the direction of II, change magnetic field force, the magnet that wherein produces magnetic field is electro-magnet;
III, change the size and/or the direction of magnetic field force and make said container and the relative movement of said magnetic field, the magnet that wherein produces magnetic field is electro-magnet.
3. method according to claim 1 and 2 is characterized in that: said to make the method for said container and the relative movement of said magnetic field be following 1), 2) or 3) shown in:
1) said container is taken exercises between two opposite rows magnet;
2) said container is around said magnet movement;
3) said magnet moves around said container.
4. according to claim 1 or 2 or 3 described methods, it is characterized in that: said 1), 2) or 3) in, said motion is circumferential motion, uniform motion or uniform circular motion.
5. according to arbitrary described method among the claim 1-4, it is characterized in that: the speed of said uniform circular motion is 8rpm-75rpm, is preferably 12rpm-30rpm or 15rpm-30rpm, is specially 15rpm, 19rpm, 23rpm or 30rpm.
6. according to arbitrary described method among the claim 1-5, it is characterized in that: during said container is taken exercises between two opposite rows magnet, the said container comb that several independent pipe coupling form of serving as reasons, and said comb is straight line.
7. method according to claim 6 is characterized in that: each independent pipe causes at the magnetic field force induced homogeneous of synchronization in the said comb.
8. method according to claim 7; It is characterized in that: make each independent pipe in the said comb synchronization the method that causes of magnetic field force induced homogeneous be: every row's magnet is connected in sequence by plurality of cuboid shape permanent magnet; Permanent magnet closure linearly and perpendicular to the N utmost point and the S utmost point line direction of permanent magnet self, the pole orientation of two adjacent permanent magnets is opposite; Two relative between two row's magnet permanent magnet edge lines are rectangular or square, and the magnetic pole of two relative permanent magnets is identical; Comb and two row's magnet equal keeping parallelisms, each rectangular-shaped permanent magnet equals the width of each single tube along the widest part of the tangent plane of the N utmost point of magnet and S utmost point line direction along the width of the tangent plane of himself N utmost point and S utmost point line direction; And comb is at the non-fringing effect regional movement of two row's magnet, and said fringing effect is the fringing effect that the two ends of every row's magnet produce.
9. according to arbitrary described method among the claim 1-8, it is characterized in that: said independent pipe is test tube or centrifuge tube; The said comb that is formed by several independent pipe coupling is eight platoon centrifuge tubes.
10. according to arbitrary described method among the claim 1-9, it is characterized in that: said target molecule is a biomolecules; Or said biomolecules is DNA, RNA, glucide or protein.
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| CN104928280A (en) * | 2015-06-26 | 2015-09-23 | 关节动力安达（天津）生物科技有限公司 | Rotary magnetic cell separation method |
| CN106399093A (en) * | 2016-12-02 | 2017-02-15 | 上海默里科基因科技有限公司 | Method and device for extracting biological molecules by virtue of magnetic beads |
| CN106754345A (en) * | 2016-12-02 | 2017-05-31 | 上海默里科基因科技有限公司 | Nucleic acid-extracting apparatus |
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