KR101282841B1 - Apparatus and method for isolating nucleicacids or biological materials - Google Patents

Abstract

The present invention relates to apparatus and methods for separating nucleic acids or biological materials. The apparatus of the present invention is provided with a tube seating portion 22 formed so that the seating space 24 is opened to the upper portion of the tube rack 20, the magnet space 26 is formed to be opened to the lower portion of the tube rack 20. . The seating space 24 and the magnet space 26 are formed in adjacent positions so as not to overlap each other, shaking by the vibration provided by the rack drive source 12 in the state in which the tube 40 is seated in the seating space 24. The operation is performed, and the magnet 36 enters the magnet space 26 while the tube 40 is installed in the seating space 24 and moves along the entire section of the material in the tube 40. In particular, the magnet 36 moves upward from the bottom of the tube 40 to provide a space in which the pipette can be moved to the top of the tube 40. In addition, in the present invention, since the magnetic particles are collected while reciprocating the magnet 36 a plurality of times, there is an advantage that the collection operation can be performed more quickly.

Description

Apparatus and method for isolating nucleic acids or biological materials

The present invention relates to an apparatus and method for isolating or purifying nucleic acids or biological materials from blood or cells or various biological samples.

Separation and purification of nucleic acids from blood and various other biological samples is an important starting point in many fields such as biology, biochemistry, molecular medicine, forensic medicine, diagnostic medicine and the like. Recently, a variety of polymerase chain reaction (PCR) reactions for DNA amplification have required frequent and essential steps in biological research and diagnosis, requiring the separation of nucleic acids from biological samples.

Conventional methods for the separation of nucleic acids have used harmful organic solvents such as phenol and chloroform. However, phenol and chloroform are harmful chemical solvents that can adversely affect workers.

In recent years, methods of using a substance having a property of binding to a nucleic acid have been proposed. Such materials include silica, glass fibers, anion exchange resins, and modified magnetic beads. These substances do not use harmful organic solvents and minimize the physical and biochemical degradation of nucleic acids during separation. In addition, the affixed nucleic acid is less affected by the enzyme that degrades the nucleic acid.

However, these methods still require the operator to manually transfer the solid material to another container using a pipette. This task involves the exposure of workers to latent viruses and bacteria from infected viruses and bacteria if they use infected blood or bacteria as a starting material for nucleic acid separation.

On the other hand, recently, a method of collecting a nucleic acid or biological material from a sample using a magnet is used, and generally a magnet enters a tube containing a sample and collects the nucleic acid or biological material from one side of the sample. However, when there is a large amount of sample in the tube, there is a problem in that the strength of the magnetic force transmitted to the bottom of the tube is weak and the collection is not uniform throughout.

In addition, even when automating a device for separating nucleic acid or biological material, in order to insert a magnet into the tube, a magnet must be mounted on the upper part of the tube to move, and such a structure supports and moves the pipette. There is a problem in that the configuration of the entire apparatus is complicated because of interference with the components to make.

Accordingly, an object of the present invention is to solve the conventional problems as described above, and to provide an apparatus capable of orbital shaking a sample contained in a tube and automatically performing a collection using a magnet.

Another object of the present invention is to be able to uniformly apply the magnetic force to a predetermined value or more for the entire section of the sample contained in the tube.

Another object of the present invention is to change the magnetic force value acting on the sample in the tube.

According to a feature of the present invention for achieving the above object, the present invention provides a base, a rack driving source mounted on the base to generate vibration, a magnet driving source mounted on the base to provide a driving force, and the rack A tube rack having a seating space with a seating space in which the tube containing the sample is seated and oscillated by the vibration generated from the driving source, and a magnet rack open to the other side, and installed on the base and mounted on the base of the magnet drive source. And a magnet inserted into the magnet space with a driving force to provide magnetic force to the tube in the seating space.

The seating space is opened in a row on the upper surface of the tube rack, and the magnet space is opened in a row on the lower surface, and the rows of the seating space and the magnet space are alternately formed.

The seating space is opened to the upper surface of the tube rack, the magnet space is opened to the lower surface, the seating space and the magnet space is formed alternately.

The magnet is installed on a magnet plate mounted to the magnet frame, which is detachably installed at a position corresponding to the magnet space, and the magnet frame is supported by a support guide on the base.

The tube rack is attached to and detached from the rack seat formed through the rack frame, and the rack frame is supported by a transmission member connected to the rack driving source.

According to another feature of the invention, the present invention by vibrating a tube containing a material containing a sample and a magnetic particle to bind the nucleic acid or biological material to the magnetic particle and by placing a magnet on the outer surface of the tube to the nucleic acid or biological material The first step of collecting the attached magnetic particles and removing the rest, and the magnetic particles attached to the nucleic acid or biological material from the first step is vibrated with the cleaning buffer to separate the foreign matter and place the magnet on the outer surface of the tube The second step of collecting and removing the remaining magnetic particles with nucleic acid or biological material attached thereto, and separating the nucleic acid or biological material from the magnetic particles by vibrating the magnetic particles with the nucleic acid or biological material attached in the second step with the elution buffer And do it on the outer surface of the tube By placing the net attracted magnetic particles comprises a third step of obtaining a nucleic acid or a biological material.

The magnet reciprocates a plurality of times along the entire surface of the material contained in the tube along the outer surface of the tube.

The magnet is moved from the bottom of the outer surface of the tube along the entire section of the material contained in the tube.

In the apparatus and method for separating nucleic acids or biological substances according to the present invention, the following effects can be obtained.

The apparatus of the present invention enables the sequencing and collection of the sample contained in the tube to be performed sequentially, thereby automating the process of separating the nucleic acid or biological material from the sample, so that even if the operator is not skilled, the nucleic acid or the biological The material can be separated and the operator can be free from the risk of infection.

In addition, in the present invention, since the magnet providing magnetic force to the sample is positioned along the outer surface from the bottom of the tube containing the sample along the outer surface, the magnetic force can be uniformly applied to the entire region of the sample, thereby improving the quality of the collection. .

In addition, in the present invention, since the magnet reciprocates a plurality of times along the outer surface of the tube containing the sample, there is an effect that the collection occurs more quickly while the change of magnetic force occurs in each region of the sample.

In the apparatus of the present invention, since the magnet approaches the tube from the lower side of the tube, the upper space of the tube becomes a space accessible to other components other than the magnet, so that it is easier to design the configuration of the entire apparatus. There is also an effect.

1 is a perspective view showing a preferred embodiment of the device for separating nucleic acid or biological material according to the present invention.
Figure 2 is a side view showing the configuration of an embodiment of the present invention.
Figure 3 is a top perspective view showing the upper configuration of the tube rack constituting an embodiment of the present invention.
Figure 4 is a bottom perspective view showing a lower configuration of the tube rack constituting an embodiment of the present invention.
5 is a cross-sectional view taken along line 5-5 of FIG. 3.
6 is a sectional view taken along line 6-6 of FIG.
Figure 7 is a working state showing the state that the magnet is located in the magnet space in the embodiment of the present invention.

Hereinafter, preferred embodiments of the apparatus and method for separating nucleic acids or biological substances according to the present invention will be described in detail with reference to the accompanying drawings.

The present invention relates to a device and a method for separating nucleic acid or biological material from a sample, in particular to obtain a desired material from the sample by using a magnetic force of the magnet. To this end, the sample containing the magnetic particles is shaken for a predetermined time, and then separated by applying a method in which a magnet is approached to the sample so that nucleic acids or biological substances attached to the magnetic particles are concentrated on one side in the sample.

As shown in the figures, the apparatus of the present invention is provided with a rack drive source 12 and a magnet drive source 14 on the base 10. In the illustrated embodiment, there are these drive sources 12, 14 across the bottom and top of the base 10. However, the driving sources 12 and 14 may be installed at both the upper and lower portions of the base 10. The base 10 is one piece, but may be mounted on the base 10 in which the rack drive source 12 and the magnet drive source 14 are separated. In addition, the base 10 may be supported on the ground or a table through a separate support device.

The transmission member 16 is installed in the upper portion of the base 10 is connected to the rack drive source (12). The transmission member 16 serves to receive and transmit the vibration generated from the rack driving source 12. The transfer member 16 is made in the shape of a plate in this embodiment. The transmission member 16 is installed to face each other at a predetermined interval between the base 10. The transfer member 16 may have a predetermined interval between the base 10 and the transfer member 16 because the transfer member 16 is directly installed in the rack driving source 12. The support 16 ′ is provided on the transfer member 16. The support 16 'is provided to stand upright at both ends of the transfer member 16 in this embodiment. The support 16 ′ serves to support the rack frame 18 which will be described below as part of the transfer member 16.

The rack frame 18 is supported by the support 16 ′ of the transfer member 16. The rack frame 18 receives the vibration of the rack drive source 12 through the transmission member 16. The rack seat portion 18 'is formed in the rack frame 18. The rack seating portion 18 ′ is formed by penetrating the rack frame 18 up and down, and the tube rack 20 to be described below is located. A seating end (not shown) is formed along an inner surface edge of the rack seating portion 18 ′ so that the bottom edge of the tube rack 20 is hooked and supported.

The tube rack 20 is positioned at the rack seating portion 18 ′ of the rack frame 18. The tube rack 20 is installed so that the upper surface is exposed to the upper portion of the rack frame 18, the lower surface is exposed to the lower portion. The tube rack 20 has a substantially rectangular plate shape in this embodiment.

The tube rack 20 is formed to protrude from the tube seating portion 22 to be exposed to the upper surface. The tube seating portion 22 is arranged in a row in a plurality of vertical lines. A seating space 24 is formed in the tube seating part 22. In this embodiment, the cross section of the seating space 24 is circular. However, the cross section of the seating space 24 corresponds to the cross sectional shape of the tube 40 and may have various shapes. The tube 40 is seated in the seating space 24. The seating space 24 is formed to almost the bottom of the tube rack (20). In the seating space 24, a tube 40 containing a sample is seated.

A magnet space 26 is formed to open to the bottom surface of the tube rack 20. The magnet space 26 has a circular cross section. The magnet space 26 is located between the seating space 24 is formed so as not to overlap with the seating space 24. Accordingly, the magnet space 26 and the inner surface of the seating space 24 have a similar thickness as a whole, and the magnet spaces 26 are also arranged in rows vertically and horizontally.

For reference, the shape of the magnet space 26 has a circular cross section in this embodiment, but may be in various forms. That is, the magnet space 26 may be made in various ways, such as a donut shape surrounding the seating space 24 or a half of the donut shape.

On the other hand, a plurality of movement guides 30 are installed upright on the base 10. The movement guide 30 serves to guide the movement of the magnet frame 32 that is moved by the magnet drive source 14. The magnet frame 32 is located between the top of the transfer member 16 and the bottom of the rack frame 18. In addition, the movement guide 30 is in a position not to interfere with the transfer member 16. The movement guide 30 is made in the shape of a rod in this embodiment, and has a length enough to guide the lifting stroke of the magnet frame 32.

The magnet frame 32 is supported by the drive shaft 14 ′ of the magnet drive source 14. The drive shaft 14 'of the magnet drive source 14 has a threaded portion formed on an outer surface thereof, and a threaded portion is formed on an inner surface of a through hole of the magnet frame 32 through which the drive shaft 14' passes. The magnet frame 32 is moved by the rotation.

The magnet plate 34 is detachably installed on the magnet frame 32. The magnet plate 34 has a plate seating groove 34 ', and a plurality of magnets 36 are arranged in a row. The magnet 36 may have a shape corresponding to that of the magnet space 26 so as to enter the magnet space 26 and provide the magnetic force uniformly to the tube 40. In this embodiment, the cross section is circular. The magnet 36 is replaceably installed on the magnet plate 34.

  Hereinafter, the separation of nucleic acids or biological materials using the apparatus and method for separating nucleic acids or biological materials according to the present invention as described above will be described.

First, the operation of the apparatus of the present invention will be described. The tube rack 20 mounted on the rack frame 18 is driven by the rack driving source 12 to vibrate. That is, the vibration generated from the rack drive source 12 is transmitted to the rack frame 18 through the transmission member 16. Therefore, the tube rack 20 seated on the rack seating portion 18 ′ of the rack frame 18 is vibrated by the rack frame 18.

Such a vibration vibrates the tube 40 seated on the tube seat 22 of the tube rack 20. Sample 40, a magnetic particle, a buffer, etc. are selectively contained in the tube 40. As the tube 40 vibrates, nucleic acids or biological substances in the sample are combined with the magnetic particles or separated from the magnetic particles.

On the other hand, the magnet 36 enters the magnet space 26 of the tube rack 20 to reciprocate. After the shaking process for the tube 40 is finished, the magnet frame 32 is moved toward the rack frame 18 by the operation of the magnet driving source 14. This is achieved by power being transferred to the magnet frame 32 by the drive shaft 14 'of the magnet drive source 14.

When the magnet frame 32 moves toward the rack frame 18, the magnet 36 enters the magnet space 26. The state in which the magnet 36 enters the magnet space 26 is shown in FIG. 7. The extent to which the magnet 36 enters the magnet space 36 is controlled by the magnet drive source 14. To this end, the magnet driving source 14 may use a step motor or a linear motor.

The magnet 36 moves in and out of the magnet space 36 to exert a magnetic force on the tube 40 contained in the adjacent seating space 24. The magnetic force causes the magnetic particles in the tube 40 to collect on the inner surface of the tube 40. When the magnetic particles are collected on the inner surface of the tube 40 in this way, the remaining material contained in the tube 40 may be removed using a pipette.

The process of isolating nucleic acid or biological material using the device of the present invention will now be described.

In order to finally separate a desired substance from a sample, that is, a biological substance such as a nucleic acid or a protein, first, a sample (for example, blood), magnetic particles (magnetic beads) and a binder are introduced into the tube 40. As an example of the binder, guanidine thiocyanate is used. The guanidine buffer serves to destroy cells in the sample and to attach nucleic acids or biological materials to the magnetic particles. The tube 40 is placed in a seating space 24 formed in the tube seating portion 22 of the tube rack 20. Of course, the tube 40 is seated on each of the tube seats 22.

When the rack drive source 12 is operated while the tube 40 is seated on the tube seat 22, vibration generated by the rack drive source 12 is transmitted to the rack frame through the transfer member 16. It is delivered to (18), the tube rack 20 vibrates to shake the operation. In this process, the magnetic particles and the nucleic acid or biological material in the sample are bound by a binder.

After shaking for a predetermined time, the rack drive source 12 is stopped and the magnet drive source 14 is driven. The magnet frame 32 is moved by the magnet drive source 14. The magnet frame 32 is moved to allow the magnet 36 to enter the magnet hole 26. When the magnet 36 enters the magnet space 26, the magnetic force affects the tube 40 in the adjacent seating space 24. Thus, magnetic particles in the tube 40 are collected toward the magnet 36.

At this time, the magnet 36, as shown in Figure 7, so that the end portion is to go to a position corresponding to the upper limit of the sample so as to exert a magnetic force on the entire sample in the tube (40). This is done by setting the movement stroke of the magnet 36 to the controller in accordance with the amount of the sample at the time of operation.

In addition, although the magnet 36 may stop in the magnet space 26 to provide magnetic force, it is faster to collect the magnetic particles by allowing the magnet 36 to move within a range corresponding to the sample. Let's do it. This is due to the change of the magnetic force due to the movement of the magnet 36.

When the magnetic particles are collected on the inner surface of the tube 40 by the magnet 36, the remaining substances in the tube 40 are removed while the magnet 36 is placed in the magnet space 26.

After removing the remainder of the magnetic particles bound to the biological material from the tube 40, the magnet 36 is driven from the magnet drive source 14 to be removed from the magnet space 26.

Next, the washing buffer is injected into the tube 40, and the rack driving source 12 is driven again to perform the shaking operation. Alcohol may be used as the cleaning buffer. This time is to remove the foreign matter adhering to the magnetic particles, that is, remaining in the magnetic particles or the tube 40 among the samples except biological material.

After the foreign matter is separated from the magnetic particles by the washing buffer using the cleaning buffer, the magnet driving source 14 is driven to allow the magnet 36 to enter the magnet space 26. Thus, the magnetic particles gather back to the inner surface of the tube 40. This operation is performed by repeatedly moving the magnet 36 along the range of material filled in the tube 40 as described above.

Then, the washing buffer is removed while the magnet 36 is fixed in the magnet space 26. This operation is repeated until the foreign matter in the tube 40 is removed to a certain level or more.

Finally, a shaking operation and a collection operation are performed to separate biological materials from the magnetic particles. That is, the introduction buffer is introduced into the tube 40. The elution buffer serves to separate biological materials from the magnetic particles, and for example, water or TE buffer may be used.

The rack driving source 12 is driven again while the elution buffer is inserted into the tube 40 to shake the biological material from the magnetic particles. After the shaking operation is performed by driving the rack drive source 12 for a predetermined time or more, the magnet drive source 14 is driven to collect the magnetic particles by the magnetic force of the magnet 36 to the inner surface of the tube 40. This operation is also done by moving the magnet (36).

In the state in which the biological material is separated from the magnetic particle and mixed with the illu- tion buffer, the magnet 36 is fixed to the magnet space 26, and the illution buffer containing the biological material is extracted and sent to the next work process. .

The scope of the present invention is not limited to the embodiments described above, but may be defined by the scope of the claims, and those skilled in the art may make various modifications and alterations within the scope of the claims It is self-evident.

For example, in the illustrated embodiment, the column of the magnet space 26 is formed in the opposite direction in which the seating space 24 is opened between the row and the row of the seating space 24 formed in the tube rack 20. It is. However, the seating space 24 and the magnet space 26 may be alternately formed so that the magnet space 26 surrounds the seating space 24. That is, the opening directions of the seating space 24 and the magnet space 26 are the same as the illustrated embodiment, but they may be alternately formed in the longitudinal and transverse directions. In addition, the arrangement of the seating space 24 and the magnet space 26 may be variously made to increase the collection performance of the magnetic particles.

10: base 12: rack drive
14: magnet drive 16: transmission member
16 ': support 18: rack frame
20: tube rack 22: tube seat
24: Seating space 26: Magnet space
30: support guide 32: magnet frame
34: magnet plate 36: magnet

Claims (8)

Bass,
A rack driving source mounted to the base to generate vibration;
A magnet driving source mounted to the base to provide a driving force;
The tube rack is formed so that the tube seat is opened to one side and the magnet space is opened to the other side with a seating space that is vibrated by the vibration generated from the rack drive source, the tube containing the sample;
And a magnet installed on the base and inserted into the magnet space by the driving force of the magnet driving source to provide a magnetic force to a tube in the seating space.
The nucleic acid or biological material of claim 1, wherein the seating spaces are opened in a row on the upper surface of the tube rack, and the magnet spaces are opened in a row on the lower surface, and the columns of the seating spaces and the magnet spaces are alternately formed. To separate the device.
The apparatus of claim 1, wherein the seating space is opened to an upper surface of the tube rack, and the magnet space is opened to a lower surface, wherein the seating space and the magnet space are alternately formed.
The method of claim 1, wherein the magnet is installed on a magnet plate mounted to the magnet frame, is detachably installed in a position corresponding to the magnet space, the magnet frame is a nucleic acid supported by a support guide on the base or Device for separating biological material.
The apparatus of claim 1, wherein the tube rack is detached and seated in a rack seat formed through the rack frame, and the rack frame separates the nucleic acid or biological material supported by the transfer member connected to the rack driving source.
By vibrating the tube containing the sample and the material containing the magnetic particles, the nucleic acid or biological material is coupled to the magnetic particle, and the magnet is placed on the outer surface of the tube to collect the magnetic particles to which the nucleic acid or biological material is attached and remove the rest. The first step,
The magnetic particle attached to the nucleic acid or biological material attached in the first step is vibrated with the cleaning buffer to separate the foreign material, and the magnet is placed on the outer surface of the tube to collect the magnetic particles to which the nucleic acid or biological material is attached and remove the rest. Step two,
The magnetic particles with the nucleic acid or biological material attached in the second step are vibrated with the illumination buffer to separate the nucleic acid or biological material from the magnetic particles and place a magnet on the outer surface of the tube to collect the magnetic particles and obtain the nucleic acid or biological material. A method of separating a nucleic acid or biological material comprising a third step.
7. The method of claim 6, wherein the magnet separates nucleic acid or biological material reciprocating multiple times along the entire surface of the material contained in the tube along the outer surface of the tube.
8. The method of claim 7, wherein the magnet separates nucleic acid or biological material from a lower portion of the outer surface of the tube and moves along the entire section of the material contained in the tube.

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