KR101471920B1 - A Convenient Portable Cell Destructible DNA extractor - Google Patents

Abstract

The present invention relates to a cell disruption and gene extractor which is easy to operate and easy to carry, and a gene extracting process from a cell tissue, which has been possible only through experiments for a certain period of time by researchers treating various equipment and reagents in a conventional laboratory, The present invention relates to a cell disruption method and a gene extractor.
The gene crusher includes a crushing vessel for crushing a biomaterial including a reagent for cell crushing and a bead, a plug including a filter and a gene amplifying unit for extracting only genes from the crushed cells, A needle for mounting a needle on the needle mounting groove, a needle mounted on the needle mounting groove, and a cap for protecting the needle.

Description

Description of the Related Art [0002] A convenient portable cell destructible DNA extractor

The present invention relates to a cell disruption and gene extractor which is easy to operate and easy to carry, and a gene extracting process from a cell tissue, which has been possible only through experiments for a certain period of time by researchers treating various equipment and reagents in a conventional laboratory, The present invention relates to a cell disruption method and a gene extractor.

In recent years, imports of environmentally polluted and irresponsible foreign agricultural and fishery products have increased the number of fish, vegetables and vegetables contaminated by mad cow disease and radiation. In this regard, the government has provided measures through the introduction of the country of origin indication system, but until now, it has taken a long time to conduct the inspection of origin or species discrimination, and there is a problem that it can be performed only in a laboratory using professional equipment.

However, consumers are becoming more and more interested in safe food, such as being illegally converted into domestic products during distribution. Therefore, there is a demand for technology that ordinary consumers and retailers can easily distinguish between origin and species of food ingredients during the purchase process.

In general, genetic information on biomaterials, including species and origin of plants and animals, and even diseases, can be found by analyzing the genes of the biomaterials. The genetic information of the biomaterial is contained in the nucleic acid of the cell. By separating the chromosomes in the nucleic acid to obtain information about DNA (deoxyribonucleic acid), it is possible to identify various kinds of biomaterials. Thus, biomaterials that cause unknown biological phenomena can be identified.

In order to obtain information on the presence and amount of a specific nucleic acid, a process of extracting nucleic acid from the chromosome in the cell of the biomaterial including the nucleic acid is preceded. Finally, in order to analyze this, the process of amplifying the extracted DNA by the amount required for the inspection is performed again. The nucleic acid extraction process is known to use a bead. The process of amplifying the extracted nucleic acid and confirming the presence or absence of a specific nucleic acid and the presence thereof is known as a process using PCR (Polymerase Chain Reaction).

However, since these processes were carried out using professional equipment, they were all carried out only by experts with expert knowledge and skills in the laboratory. Therefore, it takes a long time to obtain the analysis result, and there has been a problem that it is difficult for the general user to easily use the apparatus by using a relatively large stationary apparatus.

Korean Patent Laid-Open Publication No. 10-2010-0051494 relates to a portable device and method for real-time gene extraction of animal tissues, which comprises sampling means for sampling animal tissue cells on the surface of a drill, animal tissue cells A first tube container for dissolving the solution in the solution; driving means for rotating the sampling means to drive the drill into the first tube container; Centrifugal separating means for removing foreign substances from the solution in which the cells are dissolved, and DNA filter means for collecting the DNA by purifying the foreign substance-removed solution. However, as shown in FIGS. 1 and 2 of the prior patent documents, it is too complicated for a general consumer to use and is also referred to as a portable device, but has a configuration and size that are practically impossible to carry, Still difficult to do.

In general, the process of disrupting the cells and detecting the genes from them is a complex process that requires specialized equipment and has been done only by experts at a limited area of the laboratory. Therefore, the analysis process takes a long time, There is a problem that it is difficult to use immediately when necessary.

In order to solve this problem,

First, we want to provide a cell crusher that allows consumers to quickly obtain the results by enabling physical disruption and chemical disruption to proceed at the same time so that the cell disruption process can be accelerated.

Secondly, it has a small size and a simple structure to provide a cell crusher suitable for a user to carry, thereby providing a cell crusher having a structure that can be easily carried by a consumer.

Third, the structure of the cell crusher is simple, and the operation of inducing the crushing process is simplified so that it can be carried out by anyone. Thus, the cell crushing process for genetic testing can be performed at the purchase site, not the laboratory, So that anyone can carry out the cell disruption process at a desired time.

Fourthly, there is a ultimate goal of eliminating the anxiety about the food by providing the cell disruption method and the gene extractor which can directly discriminate the origin and diseases of the concentrated aquatic products in the field.

In order to achieve the above object,

In order to accelerate the cell disruption at high speed, the present invention allows simultaneous physical disruption and chemical disruption. That is, a bead made of ceramic, glass, polymer, metal, or the like is placed in a crushing vessel of a cell crusher, and a reagent for cell crushing is charged on the one hand to repeatedly apply force to the crushing vessel, So that the crushing and the chemical crushing by the reagent can proceed at the same time.

Also, the crushing vessel is made of a material having elasticity and flexibility so that the crushing vessel can be repetitively compressed and returned to its original state by this repetitive force.

On the other hand, the present invention is for the rapid detection of a gene. After the cell disruption, it is necessary to filter only the gene and to inject it into the gene amplifier. For this purpose, the plug portion of the cell crusher is separated from cell clusters or beads And a filter for extracting only genes.

Also, a gene amplification unit for mixing the extracted gene with the gene amplification reagent is formed inside the gene amplification unit so that the mixed solution containing the gene amplification reagent and the gene can be introduced into the gene amplification unit.

The present invention enables physical disruption and chemical disruption to proceed at the same time, thereby rapidly advancing the disintegration process of cells, and thus has a first effect that enables consumers to obtain results quickly.

In addition, the present invention provides a cell crushing and gene extractor suitable for carrying the user, which is made up of a small size and a simple structure, so that the cell can be easily carried by the consumer and can be directly subjected to cell disruption and gene extraction The second effect is achieved.

As a result, by providing a cell disruption and gene extractor capable of directly discriminating the origin and disease state of a concentrated aquatic product in the field, it has the ultimate effect of relieving the anxiety about the food of consumers.

1 is a configuration diagram of a cell crusher according to the present invention.
Fig. 2 is an internal configuration diagram of a cap of a cell crusher.
3 is a view showing a process of disrupting cells using a cell crusher.
FIG. 4 is a view showing a process of transferring the cell disruption reagent from a cell disruption reagent container to a cell disruptor.
FIG. 5 is a flowchart showing a step-by-step method of disrupting cells.
FIG. 6 is a flowchart showing a stepwise method of transferring an extracted gene to a gene amplifier after cells are disrupted.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made to solve the above-mentioned problems, and an object of the present invention is to provide a device for extracting genes from a cell which is simple and easy to carry and can be carried by a user.

One embodiment of the cell crusher of the present invention is shown in FIG.

FIG. 1 is a schematic view showing the structure of a cell crusher 100 according to the present invention. The cell crusher 100 according to the present invention includes a crushing vessel 110 having an entrance 114 formed at one side thereof for containing and crushing biomaterials, And a protrusion 121 having a diameter smaller than that of the inlet and outlet 114 protrudes in the same direction as the inlet and outlet 114. The protrusion 121 of the protrusion 121 A needle 130 for mounting a needle 130 at a central portion thereof, a needle 130 mounted on the needle mounting groove, and a cap 140 for protecting the needle. The shape and material of the crushing container 110, the inlet / outlet 114, the protrusion 121, the cap 140, the cap 120, and the needle 130 are selected according to the purpose of the user It will be apparent to those skilled in the art that it is possible to construct it.

The crushing vessel 110 must be flexible and elastic so as to be repeatedly subjected to compression and decompression as the force is applied. For this, the crushing vessel 110 may be made of elastic polymer, metal, or other flexible material .

Particularly, the present invention relates to a method for disrupting a biological material (111) in a pulverizing container (110), which comprises a cell disrupting reagent (112) for physically disrupting a biological material (111) (Bead) 113 for crushing the beads.

The cell disrupting reagent 112 may include at least one of distilled water , proteolytic enzyme, xylene, acetone, chloroform, and alcohol, The bead 113 may be made of glass, ceramics, metal, or polymer. The shape of the bead 113 can be selected according to the purpose of the user, but it is preferable to form the bead 113 in the shape of a small sphere having a diameter of 100 μm or less Crushing and the like.

The coupling between the inlet and outlet 114 of the crushing container 110 and the plug 120 may be such that the spiral protrusion 115 of the inlet and outlet 114 is inserted into the spiral groove of the plug 120 And the needle mounting groove of the cap projection 121 and the needle 130 can be coupled with each other by the elasticity of the groove after the needle 130 is mounted by the user's force have. However, it is obvious to those skilled in the art that such a coupling structure can be selected in accordance with the purpose of the user among the combination of existing reagents and simple containers, and is not limited to these embodiments.

The cell shredding machine of the present invention is used in a pretreatment step for detecting a gene and analyzing the gene. In order to detect a gene, a cell is disrupted, and a gene amplification process for amplifying an extracted gene after impurities other than the gene is removed is performed In order to do this, the entire process can be simply proceeded by extracting only the gene before transferring the gene to the gene amplifier and mixing it with the amplification reagent.

Accordingly, the present invention forms a structure for the cap portion, and FIG. 2 is an internal configuration view of the cap 120 of the cell crusher 100 for this purpose. The plug 120 includes a filter 122 for passing only genes therein and a gene amplification reagent for amplifying the gene that has passed through the filter 122 and is located at the protrusion 121 of the plug 120 And a gene amplifying unit 123 having the same.

The filter 122 may be a polypropylene, a polystyrene, a poly (vinylidene chloride), or the like, which can effectively extract genes only from the cell masses from which the gene has been removed, And at least one polymer material filter such as polyester, polyethylene terephthalate (PET), cellulose, polyvinylidene fluoride (PVDF), etc., The amplification reagent of the amplification reaction unit 123 may include at least one of a DNA polymerase, a nucleic acid precursor (dNTP), a primer, and a reaction buffer such as MgCl ₂ to amplify only the extracted gene .

As shown in FIG. 3, the cell disruptor 100 is used to disintegrate the cells 111 into the disintegration container 110 including the cell disintegration reagent 112 and the disintegration beads 113. The force 150 is applied to the crushing vessel 110 to remove the force 150 and the crushing vessel 110 is restored to its original state.

For this purpose, the crushing vessel 110 should be made of a material having elasticity and flexibility to be restored to its original state. During this repeated compression and restoration process, the crushing beads 113 inside the crushing vessel 110 are separated from the biomaterial 111 And at the same time, the internal scouring reagent 112 also repeatedly generates a strong flow, thereby actively contacting the biological material 111 and performing a chemical grinding process . Therefore, it is possible to simultaneously perform such physical and chemical disruption, so that the disruption process can be performed more quickly and simply.

In order to further simplify the use of the cell shredder 100 of the present invention, it is preferable that a container 200 containing a reagent for cell disruption is provided together with the cell shredder 100. The vessel 200 containing the reagent for cell disruption includes a receptacle 210 accommodating a cell disrupting reagent 112 and having an entrance formed at one side thereof and a lid 220 coupled to an entrance of the receptacle 210 And a protrusion 230 having a hole formed at a central portion thereof may be formed in a part of the lid.

Particularly, as shown in FIG. 4, the process of transferring the reagent 112 for cell disruption from the cell disruption reagent container 200 to the disintegration container 110 of the cell disruptor 100 includes the steps of: So that the protrusion 230 is inserted into the inlet / outlet 114 of the crushing container. For this, the protrusion 230 of the cell disruption reagent container 200 should be made smaller than the size of the inlet / outlet 114 of the crushing container 110, and through this process, It is possible to prevent water leakage, contamination, and the like.

5, the cell shredder 100 and the cell shredding reagent container 200 are prepared, and the cell shredder 100 (FIG. 5) And then the cell crushing reagent container 200 is inverted and the cell crusher 100 is used for cell crushing the cells for destruction of cells Transfer the reagent 112. Thereafter, the cell-shredding reagent container 200 is removed, the cap 120 of the cell crusher 100 is closed, and then the cap 120 is closed. (150) is pressed to compress and then release the force to restore it.

By this repetitive compressive force, the crushing beads 113 are brought into contact with and rubbed against the biomaterial 111 with a constant force and speed, resulting in physical crushing. At the same time, the crushing reagent 112 inside A strong flow is repeatedly generated, and then the biological material 111 is actively contacted and subjected to a chemical grinding process, whereby a rapid grinding process is performed.

The bead 113 may be made of one or more materials selected from glass, ceramics, metals, and polymers, and the cell disrupting reagent 112 may include distilled water, proteolytic enzymes, xylene, And may include at least one of acetone, chloroform, and alcohol.

The cell disruption method of the present invention is a pretreatment process for detecting a gene and analyzing the gene. In order to detect a gene, a gene amplification process for amplifying a gene extracted after impurities other than a gene is removed is performed as a follow- . Therefore, a method of extracting only genes and mixing them with an amplification reagent and transferring them to a gene amplifier is shown in FIG. 6 as another embodiment of the present invention.

6, in the method of transferring the gene extracted from the cell crusher 100 to the gene amplifier, the cell crusher 100 in a state where the cell is first broken and the gene is separated is tilted in the direction of the needle , The needle 130 of the cell crusher is mounted at the entrance of the gene amplifier and the crushed solution is passed through the filter 122 existing inside the plug 120 to extract only the gene. The gene that has passed through the filter 122 is injected into the gene amplification unit 123 again and mixed with the gene amplification reagent. Then, the gene mixed with the gene amplification reagent is injected through the needle 130 into the gene amplifier To the inlet of the second chamber.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it should be understood that various changes and modifications will be apparent to those skilled in the art. Obviously, the invention is not limited to the embodiments described above. Accordingly, the scope of protection of the present invention should be construed according to the following claims, and all technical ideas which fall within the scope of equivalence by alteration, substitution, substitution, Range. In addition, it should be clarified that some configurations of the drawings are intended to explain the configuration more clearly and are provided in an exaggerated or reduced size than the actual configuration.

100: cell crusher 110: crushing vessel
111: Biomaterial 112: Cell disruption reagent
113: bead for cell disintegration 114: inlet / outlet
115: coupling spiral projection 120: cap
121: protrusion 122: filter
123: gene amplification unit 130: needle
140: Cap 150: Power
200: Cell disruption reagent container 210:
220: lid 230: protrusion

Claims (18)

In a portable cell shredder which is easy to operate,
A crushing container having an entrance formed at one side thereof for containing and crushing biomaterial;
A plug coupled to an inlet and an outlet of the crushing vessel and having a shape in which a protrusion having a smaller diameter than the inlet and outlet protrudes in the same direction as the inlet and outlet;
A needle mounting groove formed at a central portion of the projection of the cap to be attached with a needle;
A needle mounted on the needle mounting groove;
A cap for protecting the needle;
And,
Wherein the crushing vessel includes a cell crushing reagent for chemically crushing the biomaterial and a bead for crushing physically,
Wherein the plug includes a filter for passing only a gene therein and a gene amplification unit located at a protrusion of the plug and containing a gene amplification reagent for amplifying a gene that has passed through the filter.
The method according to claim 1,
Wherein the crushing vessel has flexibility and elasticity so that compression and restoration can be repeatedly performed as the force is applied.
delete The method according to claim 1,
Wherein the cell disruption reagent comprises at least one of distilled water, proteolytic enzyme, xylene, acetone, chloroform, and alcohol.
The method according to claim 1,
Wherein the pulverizing bead is made of at least one material selected from the group consisting of glass, ceramics, metal, and polymer.
The cell shredder according to claim 1, wherein a coupling between the inlet and outlet of the crushing container and the plug is coupled with the spiral protrusion of the inlet and outlet in a spiral groove of the plug.
The method according to claim 1,
Wherein the needle mounting groove of the cap projection and the needle are engaged by contraction due to elasticity of the groove after the needle is mounted.
delete The method according to claim 1,
The filter may be a polymeric material such as polypropylene, polystyrene, polyester, polyethylene terephthalate (PET), cellulose, polyvinylidene fluoride (PVDF) A filter, and a filter.
The cell shredder according to claim 1, wherein the gene amplification reagent comprises at least one of a DNA polymerase, a nucleic acid precursor (dNTP), a primer, and a reaction buffer of MgCl ₂ . .
delete delete delete In a method of disrupting cells and extracting genes,
(i) preparing the cell shredder of claim 1;
(ii) a container for cell disruption reagent, which contains a reagent for cell disruption, a receptacle having an inlet formed at one side thereof, and a lid for coupling with an entrance of the receptacle, and a protrusion formed at a central portion of the lid, ;
(iii) placing bead for grinding in the crushing vessel of the cell crusher;
(iv) immersing the biological material in the crushing vessel of the cell crusher;
(v) transferring the reagent for cell disruption from the cell disruption reagent container to the cell disruptor;
(vi) closing the cap of the cell shredder;
(vii) repeating a process of compressing and compressing the crushing vessel of the cell crusher with the plug closed to restore the cells to be crushed;
(viii) extracting genes by removing intracellular impurities from the disrupted cells;
Wherein the cell is disrupted and the gene is extracted.
15. The method of claim 14,
Wherein the bead for milling in step (iii) is made of at least one material selected from the group consisting of glass, ceramics, metal, and polymer.
15. The method of claim 14,
Wherein the reagent for cell disruption in step (v) comprises at least one of distilled water, proteolytic enzyme, xylene, acetone, chloroform, and alcohol. How to break down cells and extract genes.
delete A method for transferring a gene extracted from a cell crusher according to claim 1 to a gene amplifier,
(a) tilting the cell crusher of claim 1 in the direction of the needle, and attaching the needle of the cell crusher to the entrance of the gene amplifier;
(b) passing only the gene in the solution disrupted with the filter;
(c) injecting the gene, which has passed through the filter, into the gene amplification unit and mixing with the gene amplification reagent;
(d) transferring the gene mixed with the gene amplification reagent to the entrance of the gene amplifier through the needle;
Wherein the gene is amplified by a gene amplification method.

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