Magnetic bead extraction kit

Abstract

The utility model discloses a magnetic bead extraction kit which comprises a sample adding plate which comprises at least one group of impurity removal magnetic bead holes, a purification magnetic bead hole, a storage hole and an elution hole; the impurity removal magnetic bead hole comprises at least two reaction cavities; an impurity-removing magnetic bead solution and impurity-removing magnetic beads are accommodated in the magnetic bead solution cavity, and the impurity-removing magnetic beads are used for removing impurities from nucleic acid in a biological sample to be extracted; a purified magnetic bead solution and purified magnetic beads are accommodated in the purified magnetic bead holes, and the purified magnetic beads are used for adsorbing nucleic acid in the biological sample to be extracted. The sample adding plate is provided with the purification magnetic bead hole and the impurity removal magnetic bead hole, and is also provided with the storage hole, so that the purification magnetic bead or the impurity removal magnetic bead can be temporarily transferred and stored in the hole position for storage during use by providing an unoccupied hole position during use, and the kit can be matched to complete efficient extraction without need.

Description

TECHNICAL FIELD

[0001] The utility model belongs to molecular biology experimental equipment technical field, concretely relates to a kind of magnetic bead extraction kit. BACKGROUND

[0002] Protein is the basic substance of life, nucleic acid is the original template of expressing protein. From complex sample, various pathogens, high-quality nucleic acid is extracted in cell, is the cornerstone of transcription, translation high-quality protein. Complex sample contains rich impurities, which affects the effect of nucleic acid extraction. The conventional method for removing impurities includes centrifugation, filtration, precipitation, etc. Automatic nucleic acid extraction realizes the automatic extraction of nucleic acid in the sample through a nucleic acid extractor. The functions of the extractor mainly include system mixing, magnetic bead adsorption, cleaning and other steps, so as to realize the separation of nucleic acid and impurities. Then the nucleic acid substance is released from the eluent, and the extraction of nucleic acid is finally completed. Since it does not include centrifugation, filtration, sedimentation and other functions, only the conventional cleaning method is used to remove impurities, the quality of the extracted nucleic acid is not high, so a special centrifuge is usually required to improve the quality of the nucleic acid, which increases the cost.

[0003] Therefore, there is an urgent need for a nucleic acid extraction kit that does not require centrifugation to remove impurities and is simple to operate, to achieve efficient and high-quality nucleic acid extraction. UTILITY MODEL CONTENT

[0004] To solve the above technical problems, the utility model provides a kind of kit, by the magnetic bead of removing impurity first to the nucleic acid of the biological sample to be extracted in removing impurity, then again by the magnetic bead of purifying to the nucleic acid of the biological sample to be extracted in purifying, so as to not need centrifugation to remove impurity, can realize efficient and high-quality nucleic acid, and operation is simple, and cost is low.

[0005] The utility model provides a kind of magnetic bead extraction kit, it includes:

[0006] The sample plate includes at least one set of impurity-removing magnetic bead hole, purification magnetic bead hole, storage hole and elution hole;

[0007] The impurity-removing magnetic bead hole includes at least two reaction cavities completely separated by a separation film. The at least two reaction cavities include a magnetic bead solution cavity and a lysis solution cavity. The magnetic bead solution cavity contains an impurity-removing magnetic bead solution. The impurity-removing magnetic bead solution includes impurity-removing magnetic beads, which are used for removing impurities from nucleic acid in a biological sample to be extracted. The lysis solution cavity contains a lysis solution, which is used for lysing cell membranes to release nucleic acid. The separation film is easy to break.

[0008] The purification magnetic bead hole contains a purification magnetic bead solution, which includes purification magnetic beads used for adsorbing nucleic acid in the biological sample to be extracted.

[0009] The storage hole is used for temporarily storing the purification magnetic beads or the impurity-removing magnetic beads in use; and

[0010] The elution hole contains an elution solution, which is used for separating the nucleic acid from the purification magnetic beads.

[0011] Preferably, when the nucleic acid is extracted, the storage hole is used for temporarily storing the purification magnetic beads, and the purification magnetic bead hole is also used for temporarily storing the impurity-removing magnetic beads after adsorbing impurities; and / or each group further comprises a washing hole for storing a washing solution, or

[0012] The storage hole also contains a washing solution;

[0013] The washing solution is used for washing the purification magnetic beads after adsorbing the nucleic acid.

[0014] Preferably, the outer surface of the magnetic microsphere body of the impurity-removing magnetic bead has at least one branched polymer with a functional group.

[0015] Preferably, the depth of the impurity-removing magnetic bead hole, the purification magnetic bead hole, the storage hole and the elution hole is 20-50 mm, and / or the cross section of each hole is square.

[0016] Preferably, the sample loading plate sequentially distributes at least one group of the impurity-removing magnetic bead hole, the purification magnetic bead hole, the storage hole and the elution hole in the row or column direction.

[0017] Preferably, for a group of impurity-removing magnetic bead hole, the purification magnetic bead hole, the storage hole and the elution hole, the length of the impurity-removing magnetic bead hole is at least twice the length of the purification magnetic bead hole, and the length of the purification magnetic bead hole is the same as the length of the storage hole and the elution hole.

[0018] Preferably, the magnetic bead solution cavity of the impurity-removing magnetic bead hole also contains a neutralizing solution; or

[0019] The impurity-removing magnetic bead hole further comprises a neutralizing solution cavity, which is located between the magnetic bead solution cavity and the lysis solution cavity, wherein the neutralizing solution cavity contains a neutralizing solution.

[0020] Preferably, the separation membrane horizontally separates the magnetic bead solution cavity and the lysis solution cavity, and the magnetic bead solution cavity is located below the lysis solution cavity.

[0021] Preferably, the separation membrane is formed by pouring and solidifying after heating the material to a molten state, and the material of the separation membrane is selected from paraffin.

[0022] Preferably, a film sealing layer is further included for sealing the sample plate.

[0023] The utility model discloses the beneficial effects are as follows:

[0024] The magnetic bead kit provided by the utility model, through setting purification magnetic bead holes and impurity removing magnetic bead holes on the sample plate, the impurity removing magnetic bead holes include at least two reaction cavities separated by a separation film, and storage holes are further arranged, through the arrangement of the storage holes, the free hole position can be provided in use, so that the purification magnetic bead or the impurity removing magnetic bead can be temporarily transferred and stored in the hole position in use, so that the impurity removing magnetic bead in the mixed solution obtained by breaking the separation film between the magnetic bead solution cavity and the lysis liquid cavity of the impurity removing magnetic bead hole can be transferred to the free hole position, so that the kit with the separation between the magnetic bead solution cavity and the lysis liquid cavity can be suitable for increasing the impurity removing method, and the high-efficiency and high-quality automatic nucleic acid extractor operation of nucleic acid can be realized without centrifugation and impurity removal, further meeting the needs of automation and high throughput.

[0025] And, since the storage holes can also be used for pre-packaged storage of cleaning liquid, it is not necessary to additionally arrange cleaning holes, so that the purification magnetic bead after extraction of nucleic acid can be cleaned, and under the premise of not increasing the excess hole position, the adverse effects caused by long-term contact of the magnetic bead with the cleaning liquid can be avoided. BRIEF DESCRIPTION OF DRAWINGS

[0026] In order to more clearly illustrate the technical scheme of the utility model, the following will be briefly introduced the drawings needed to be used in the embodiment or prior art description, obviously, the drawings in the following description are only some embodiments of the utility model, and for those skilled in the art, other drawings can also be obtained according to these drawings without creating labor.

[0027] Figure 1 It is the structure diagram of the 96-hole sample plate in the prior art;

[0028] Figure 2 It is the top view of the sample plate of one embodiment of the utility model;

[0029] Figure 3 It is the sectional view of the sample plate of one embodiment of the utility model;

[0030] Figure 4 It is the sectional view of the sample plate of another embodiment of the utility model;

[0031] Figure 5 It is the sectional view of the sample plate of another embodiment of the utility model;

[0032] Figure 6A sectional view of the sample adding plate according to another embodiment of the present application;

[0033] In the drawings:

[0034] 100: 96-hole sample adding plate, 101: standard hole

[0035] 200: sample adding plate

[0036] 210: impurity-removing magnetic bead hole, 220: purification magnetic bead hole, 230: storage hole, 240: elution hole, 250: washing hole

[0037] 211: magnetic bead solution cavity, 212: lysis solution cavity, 213: neutralization solution cavity, 214: suspension cavity DETAILED DESCRIPTION

[0038] The present application provides a magnetic bead kit, in order to make the purpose, technical scheme and effect of the present application more clear and definite, the following will be further described in detail with reference to the drawings and examples. It should be understood that the specific embodiments described herein are only used to explain the present application, and are not used to limit the present application.

[0039] As Figure 1 shown, it is a structural schematic diagram of a 96-hole sample adding plate 100 commonly used in the art, in the present application, in order to facilitate description, it is specified that one hole in the 96-hole sample adding plate 100 is a standard hole 101.

[0040] In the present application, the specific structure of the impurity-removing magnetic bead is the magnetic microsphere C in the patent CN112111042B, and the contents related to the magnetic microsphere C in the patent CN112111042B are all introduced into the present application.

[0041] In the following embodiments, the purification magnetic beads (conventional magnetic beads) are all directly modified with silicon hydroxyl or silicon carboxyl on the outer surface of the magnetic microsphere body.

[0042] The present application provides a magnetic bead kit, comprising a sample adding plate, and various solutions configured according to specific application scenarios are pre-stored on the sample adding plate. Specifically, as Figure 2 and Figure 3 shown, it is a top view of the sample adding plate 200, comprising at least one set of impurity-removing magnetic bead hole 210, purification magnetic bead hole 220, storage hole 230 and elution hole 240.

[0043] The decontamination magnetic bead hole 210 includes at least two reaction cavities completely separated by a separation film, and the at least two reaction cavities include a magnetic bead solution cavity 211 and a lysis solution cavity 212; the decontamination magnetic bead solution is accommodated in the magnetic bead solution cavity 211, and the decontamination magnetic bead solution includes decontamination magnetic beads for decontaminating nucleic acids in a biological sample to be extracted; the lysis solution is accommodated in the lysis solution cavity 212, and the lysis solution is used to break the cell membrane to release nucleic acids; wherein the separation film is easy to break.

[0044] The purification magnetic bead hole 220 accommodates a purification magnetic bead solution, and the purification magnetic bead solution includes purification magnetic beads for adsorbing nucleic acids in a biological sample to be extracted.

[0045] The storage hole 230 is used to temporarily store the purification magnetic beads or the decontamination magnetic beads during use. Through the arrangement of the storage hole 230, the spare hole position can be provided to temporarily store the transferred purification magnetic beads or decontamination magnetic beads, so as to provide extra hole positions for extraction, and the decontamination magnetic beads in the solution obtained after the direct separation film between the magnetic bead solution cavity 211 and the lysis solution cavity 212 is broken can be transferred to the extra hole positions for storage, thereby being capable of cooperating to realize increased decontamination operations to improve the purification effect.

[0046] The elution hole 240 accommodates an elution solution, and the elution solution is used to separate nucleic acids from the purification magnetic beads.

[0047] In some embodiments, in order to more cleanly decontaminate nucleic acids in a biological sample to be extracted, the outer surface of the magnetic microsphere body of the decontamination magnetic bead has at least one branched polymer, and the branched chain contains a functional group.

[0048] In some embodiments, the separation film completely separates the magnetic bead solution cavity 211 and the lysis solution cavity 212 horizontally, and the lysis solution cavity 212 and the magnetic bead solution cavity 211 are arranged in sequence from top to bottom, that is, the magnetic bead solution cavity 211 is located below the lysis solution cavity 212; at this time, the magnetic bead solution cavity 211 and the lysis solution cavity 212 are arranged in the horizontal direction, and the magnetic bead solution cavity 211 can be located on the left side of the lysis solution cavity 212 or on the right side of the lysis solution cavity 212, and the specific arrangement order is arranged according to the specific application scene. Of course, in some embodiments, the separation film can also vertically separate the magnetic bead solution cavity 211 and the lysis solution cavity 212. Since the magnetic bead solution cavity and the lysis solution cavity are separated by the separation film which is easy to break, it can be suitable for the operation of an automatic nucleic acid extractor, further meeting the needs of automation and high throughput. In addition, "horizontal" here refers to the horizontal or vertical placement of the separation film, that is, the separation film is placed as straight as possible without inclination. "In sequence" in this paper refers to the order of the above-mentioned order, which is convenient for the order required by the process to be completed.

[0049] In the embodiment in which the magnetic bead solution cavity 211 and the lysis solution cavity 212 are physically separated by the separation film, the neutralization solution can also be accommodated in the magnetic bead solution cavity 211 of the impurity removal magnetic bead hole 210, for neutralizing the uniformly lysed sample solution, so that the biological molecules to be purified can be more easily adsorbed. At this time, the magnetic bead solution cavity 211 of the impurity removal magnetic bead hole 210 not only accommodates the impurity removal magnetic beads and the impurity removal magnetic bead solution, but also accommodates the neutralization solution, that is, the magnetic bead solution cavity 211 of the impurity removal magnetic bead hole 210 accommodates the mixed solution formed by the impurity removal magnetic beads, the impurity removal magnetic bead solution and the neutralization solution.

[0050] In still other embodiments, referring to Figure 4 , the impurity removal magnetic bead hole 210 can also include a neutralization solution cavity 213, which is located between the magnetic bead solution cavity 211 and the lysis solution cavity 212. The lysis solution cavity 212, the neutralization solution cavity 213 and the magnetic bead solution cavity 211 are arranged in order from top to bottom or from left to right, which is convenient for operation. At this time, the impurity removal magnetic bead hole 210 is completely separated from the three reaction cavities: the lysis solution cavity 212, the neutralization solution cavity 213 and the magnetic bead solution cavity 211 by two layers of separation films. Preferably, the first layer of separation film is used to completely separate the lysis solution cavity 212 and the neutralization solution cavity 213 horizontally and vertically, and of course the first layer of separation film can also be used to completely separate the lysis solution cavity 212 and the neutralization solution cavity 213 vertically; the second layer of separation film is used to completely separate the neutralization solution cavity 213 and the magnetic bead solution cavity 211 horizontally and vertically, and of course the second layer of separation film can also be used to completely separate the neutralization solution cavity 213 and the magnetic bead solution cavity 211 vertically. Among them:

[0051] The lysis solution cavity 212 accommodates a lysis solution for lysing the cell membrane in the sample to release nucleic acids;

[0052] The neutralization solution cavity 213 accommodates a neutralization solution for neutralizing the uniformly lysed sample solution, so that the biological molecules to be purified can be more easily adsorbed;

[0053] The magnetic bead solution cavity 211 accommodates an impurity removal magnetic bead solution, which includes impurity removal magnetic beads for removing impurities in the biological sample to be extracted.

[0054] It should be noted that the separation film is made of an inert material that is easy to break, and the thickness of the separation film is adjusted to be broken once, preferably, the separation film is cast and cured by heating the material with a solidification temperature higher than the ambient temperature to a molten state, for example, the material of the separation film can be selected as paraffin wax, because paraffin wax has stable chemical properties and is not easy to chemically react with the separated solution; lighter than water, after being broken, it automatically floats up, and does not affect the components in the solution; and it is also flexible and easy to break. In this paper, "inert" means that it will not chemically react with the reagent solution in contact, that is, it will not affect the extraction and purification. In addition, the "film" in the separation film has no special meaning, that is, it is not limited to the thickness or material of the film as usually described, but any film that can be easily broken and separated in use.

[0055] In some embodiments, the impurity removal magnetic bead hole 210, the purification magnetic bead hole 220, the storage hole 230, and the elution hole are deep holes, and the depth is 20mm-50mm, preferably 35mm. Because the capacity of the deep hole is large, it can accommodate solutions of different dosages, thereby adapting to different application scenarios.

[0056] In some embodiments, the cross section of the impurity removal magnetic bead hole 210, the purification magnetic bead hole 220, the storage hole 230, and the elution hole 240 is selected but not limited to square. The square cross section helps to arrange the holes adjacent to each other, thereby reducing the overall length of the sample plate.

[0057] In specific implementation, the sample plate can have at least one set of impurity removal magnetic bead holes 210, purification magnetic bead holes 220, storage holes 230, and elution holes 240 distributed in the row direction in sequence; of course, the sample plate can also have at least one set of impurity removal magnetic bead holes 210, purification magnetic bead holes 220, storage holes 230, and elution holes 240 distributed in the column direction in sequence. That is, in specific implementation, the distribution direction of each group of holes on the sample plate can be selected according to different application scenarios.

[0058] Because the impurity removal magnetic bead hole 210 not only needs to accommodate impurity removal magnetic beads, magnetic bead solution, and lysis solution, but also needs to accommodate sample solution during nucleic acid extraction, the impurity removal magnetic bead hole 210 is usually a large-capacity hole. In order to facilitate modification on the existing reagent box, for a set of impurity removal magnetic bead holes 210, purification magnetic bead holes 220, storage holes 230, and elution holes 240, the length of the impurity removal magnetic bead hole 210 is set to be at least twice, preferably twice or three times, the length of the purification magnetic bead hole 220, and the lengths of the purification magnetic bead hole 220, the storage hole 230, and the elution hole 240 are the same. In some embodiments, the lengths of the purification magnetic bead hole 220, the storage hole 230, and the elution hole 240 can be the same as the length of the standard hole 101, and at this time, the length of the impurity removal magnetic bead hole 210 is at least twice the length of the standard hole 101.

[0059] In one embodiment, referring to Figure 5 In addition, a washing hole 250 for containing a washing solution for washing the purified magnetic beads after adsorbing the nucleic acid is arranged between the storage hole 230 and the elution hole 240. In some embodiments, multiple washing holes 250 can be arranged to wash the purified magnetic beads after adsorbing the nucleic acid multiple times, so as to ensure that the extracted nucleic acid is purer.

[0060] In some embodiments, the storage hole 230 can be used to temporarily store the purified magnetic beads in use, specifically, the storage hole 230 is used to temporarily store the purified magnetic beads in the process of extracting the nucleic acid, at this time, in one preferred embodiment, the storage hole 230 is also used to pre-package the washing solution, which can wash the purified magnetic beads after extracting the nucleic acid, so that the washing function can be provided without additionally arranging the washing hole 250, and the purified magnetic beads after extracting the nucleic acid can be washed, and at the same time, without increasing the extra hole position, the adverse effects caused by the long-term contact of the magnetic beads with the washing solution can be avoided: for example, in some schemes, in order to have the washing function and not increase the extra hole position, the magnetic beads for removing impurities can be stored in the washing solution, and in some schemes, the purified magnetic beads can be pre-stored in the washing solution, and the magnetic beads for removing impurities separated by the separation membrane and the storage hole 230 pre-packaged with the washing solution arranged additionally can provide the washing function while avoiding the adverse effects caused by the pre-stored magnetic beads in the washing solution.

[0061] The scheme of pre-packaging the washing solution in the storage hole 230 can also provide multiple washing functions compared with the scheme of arranging only the washing hole 250.

[0062] The specific process of extracting the nucleic acid in the biological sample when the storage hole 230 is used to temporarily store the purified magnetic beads in use will be described in detail below, taking the magnetic beads for removing impurities hole 210 being completely separated from the magnetic bead solution cavity 211 and the lysis solution cavity 212 by one layer of separation membrane as an example:

[0063] Before extracting the nucleic acid in the biological sample, the reagent kit sample plate is first pre-packaged: first, the magnetic beads for removing impurities, the magnetic beads for removing impurities solution, and the neutralizing solution are pre-packaged in the magnetic bead solution cavity 211 of the magnetic beads for removing impurities hole 210 of the sample plate 200 to form a mixed solution, when the injection amount is sufficient, then the paraffin is heated to form a liquid wax, which is dropped on the surface of the magnetic bead solution cavity, and then cooled and solidified to form a wax layer, and then the lysis solution is pre-packaged above the wax layer to complete the preparation of the magnetic beads for removing impurities hole 210 in the reagent kit; then, the purified magnetic bead solution is pre-packaged in the purified magnetic bead hole 220; then, the washing solution is pre-packaged in the storage hole 230; finally, the elution solution is pre-packaged in the elution hole 240.

[0064] After pre-dispensing the reagent box 200, the nucleic acid is extracted using the pre-dispensed reagent box, and the instrument is set to automatically extract the nucleic acid, and the specific steps are as follows:

[0065] (1) Add the biological sample liquid containing nucleic acid to be extracted into the lysis liquid cavity 212;

[0066] (2) Break the separation film layer in the impurity removal magnetic bead hole 210, so that the lysis liquid is mixed with the impurity removal magnetic beads and the mixed liquid formed by the impurity removal magnetic bead solution and the neutralization liquid, so that the impurity removal magnetic beads adsorb and remove the nucleic acid released from the sample liquid;

[0067] (3) Transfer the purification magnetic beads in the purification magnetic bead hole 220 to the storage hole 230 to temporarily store the purification magnetic beads;

[0068] (4) Transfer the impurity removal magnetic beads after adsorption and removal of impurities from the impurity removal magnetic bead hole 219 to the purification magnetic bead hole 220;

[0069] (5) Transfer the purification magnetic beads in the storage hole 230 to the impurity removal magnetic bead hole 210 to adsorb the nucleic acid;

[0070] (6) Transfer the purification magnetic beads adsorbed with the nucleic acid from the impurity removal magnetic bead hole 210 to the storage hole 230 to wash the purification magnetic beads with the washing liquid;

[0071] (7) Transfer the purification magnetic beads adsorbed with the nucleic acid from the storage hole 230 to the elution hole 240 to elute, and discard the purification magnetic beads by magnetic separation in the elution hole 240, and the separated nucleic acid is retained in the elution liquid;

[0072] (8) Release the nucleic acid from the elution liquid, and finally complete the extraction of the nucleic acid.

[0073] In the above embodiment, through the setting of the storage hole 230, the empty hole position can be provided in use, so that the purification magnetic beads can be transferred and stored in the hole position in step (3) above, so that the impurity removal magnetic beads after adsorption and removal of impurities in step (4) above are transferred to the purification magnetic bead hole for placement, so that the reagent box with separation between the magnetic bead solution cavity and the lysis liquid cavity can be appropriately increased for impurity removal extraction. This reagent box can be used for high-efficiency and high-quality automatic nucleic acid extraction instrument operation without centrifugation for impurity removal, further meeting the needs of automation and high throughput; and since the storage hole can also be used for pre-dispensing and storing the washing liquid, it is not necessary to additionally provide a washing hole, so that the purification magnetic beads after extraction of the nucleic acid can be washed, and at the same time, without increasing the extra hole position, the adverse effects caused by long-term contact of the magnetic beads with the washing liquid can be avoided.

[0074] In a more preferable embodiment, a washing hole 250 containing washing liquid can also be arranged between the storage hole 230 and the elution hole 240. After the purified magnetic beads are washed with the washing liquid in the storage hole 230 in the above step (6), the purified magnetic beads are transferred to the washing hole 250 for further washing, and then the purified magnetic beads are transferred from the storage hole 230 to the elution hole 240 for elution, separation and release of the nucleic acid, and finally the extraction of the nucleic acid is completed. Since the purified magnetic beads adsorbing the nucleic acid are washed not only by the washing liquid in the storage hole 230 but also by the washing liquid in the washing hole 250 before elution, the extracted nucleic acid can be ensured to be purer. In some embodiments, multiple washing holes 250 can be arranged to wash the purified magnetic beads adsorbing the nucleic acid multiple times, so as to ensure that the extracted nucleic acid is purer.

[0075] In yet another embodiment, the storage hole 230 can also be used to temporarily store the decontamination magnetic beads in use. Specifically, the storage hole 230 is used to temporarily store the decontamination magnetic beads in the process of extracting the nucleic acid. The following still takes the example that the decontamination magnetic bead hole 210 is completely separated from the magnetic bead solution cavity 211 and the lysis liquid cavity 212 by one layer of separation membrane, and details the specific process of extracting the nucleic acid in the biological sample when the storage hole 230 is used to temporarily store the decontamination magnetic beads in use. It should be noted that in this embodiment, the process of pre-dispensing the reagent kit sample plate can be the same as the process of pre-dispensing the reagent kit sample plate in the previous embodiment, or it can be different. The difference lies in that the storage hole 230 does not dispense the washing liquid.

[0076] After pre-dispensing the reagent kit sample plate, the nucleic acid is extracted using the pre-dispensed reagent kit, and the instrument automatic extraction program is set to extract the nucleic acid. The specific steps are as follows:

[0077] (1) adding the biological sample liquid containing the nucleic acid to be extracted into the lysis liquid cavity 212;

[0078] (2) breaking the separation membrane layer in the decontamination magnetic bead hole 210, so that the lysis liquid is mixed uniformly with the decontamination magnetic beads and the mixed liquid formed by the decontamination magnetic bead solution and the neutralization liquid, so that the decontamination magnetic beads in the decontamination magnetic bead hole adsorb and decontaminate the nucleic acid released from the sample liquid;

[0079] (3) transferring the decontamination magnetic beads after adsorption and decontamination from the decontamination magnetic bead hole 210 to the storage hole 230 for placement;

[0080] (4) transferring the purified magnetic beads from the purified magnetic bead hole 220 to the decontamination magnetic bead hole 210 for adsorption of the nucleic acid;

[0081] (5) The purified magnetic beads adsorbed with the nucleic acid are transferred from the impurity removal magnetic bead hole 210 to the elution hole 240 for elution. The purified magnetic beads are removed by magnetic separation in the elution hole 240, and the separated nucleic acid is retained in the elution solution;

[0082] (6) The nucleic acid is released from the elution solution, and the extraction of the nucleic acid is finally completed.

[0083] In the above embodiment, by providing the storage hole 230, the adsorbed impurity removal magnetic beads after impurity removal can be transferred and stored in the hole position in use, so that the method of the kit with the separation between the magnetic bead solution cavity and the lysis solution cavity can be appropriately increased for impurity removal extraction. This kit can be applied to the operation of the automatic nucleic acid extractor without centrifugal impurity removal, so as to realize the efficient and high-quality automatic nucleic acid extraction, and further meet the demand for automation and high throughput.

[0084] In a more preferred embodiment, a washing hole or a plurality of washing holes can be arranged between the storage hole 230 and the elution hole 240 to wash the purified magnetic beads adsorbed with the nucleic acid, so as to ensure that the extracted nucleic acid is more pure.

[0085] In some other embodiments, referring to Figure 6 , the impurity removal magnetic bead hole 210 can further include a suspension cavity 214. The suspension cavity 214, the lysis solution cavity 212, the neutralization solution cavity 213 and the magnetic bead solution cavity 211 are arranged in sequence from top to bottom or from left to right, which is convenient for operation. At this time, the impurity removal magnetic bead hole 210 is completely separated from the four reaction cavities by three layers of separation membranes: the suspension cavity 214, the lysis solution cavity 212, the neutralization solution cavity 213 and the magnetic bead solution cavity 211. Preferably, the first layer of separation membrane is used to completely separate the suspension cavity 214 from the lysis solution cavity 212 horizontally, and of course the first layer of separation membrane can also be used to completely separate the suspension cavity 214 from the lysis solution cavity 212 vertically; the second layer of separation membrane is used to completely separate the lysis solution cavity 212 from the neutralization solution cavity 213 horizontally, and of course the second layer of separation membrane can also be used to completely separate the lysis solution cavity 212 from the neutralization solution cavity 213 vertically; the third layer of separation membrane is used to completely separate the neutralization solution cavity 213 from the magnetic bead solution cavity 211 horizontally, and of course the third layer of separation membrane can also be used to completely separate the neutralization solution cavity 213 from the magnetic bead solution cavity 211 vertically. Among them:

[0086] The suspension cavity 214 contains a suspension, which is used to uniformly suspend the to-be-extracted substance in the sample solution in the suspension.

[0087] The lysis solution cavity 212 contains a lysis solution, which is used to lyse the cell membrane in the sample to release the nucleic acid;

[0088] The neutralization liquid cavity 213 contains neutralization liquid, which is used to neutralize the sample liquid after uniform lysis, so that the biological molecules to be purified can be more easily adsorbed;

[0089] The magnetic bead solution cavity 211 contains a decontamination magnetic bead solution, which includes decontamination magnetic beads, and the decontamination magnetic beads are used to decontaminate the nucleic acid in the biological sample to be extracted.

[0090] When pre-packaging the reagent kit sample plate of the above embodiment, first, the decontamination magnetic beads and the decontamination magnetic bead solution are injected into the magnetic bead solution cavity 211, and when the injection amount is sufficient, the paraffin is heated to form liquid wax, which is dropped onto the surface of the magnetic bead solution cavity 211, and then cooled and solidified to form a first wax layer. Then, the neutralization liquid is dispensed above the first wax layer, and when the injection amount is sufficient, the paraffin is heated again or the previously stored heated liquid wax is dropped onto the surface of the neutralization liquid cavity 213, and then cooled and solidified to form a second wax layer. Then, the lysis liquid is dispensed above the second wax layer, and when the injection amount is sufficient, the paraffin is heated again or the previously stored heated liquid wax is dropped onto the surface of the lysis liquid cavity 212, and then cooled and solidified to form a third wax layer. Then, the suspension is dispensed above the third wax layer, thereby completing the pre-packaging of the decontamination magnetic bead hole 210 in the reagent kit. After that, the specific steps of extracting nucleic acid using the pre-packaged reagent kit are similar to the above embodiment, and the difference is that after the separation membrane of the decontamination magnetic bead hole is broken, the mixed solution is different. Therefore, the specific steps of extracting nucleic acid using the pre-packaged reagent kit are not described again.

[0091] In order to facilitate the movement and transportation of the reagent kit and standardized extraction, the above sample plate can further include a sealing film layer (not shown), which is used to seal the sample plate to prevent solution leakage.

[0092] The reagent kit provided by the utility model, by setting not only the purification magnetic bead hole but also the decontamination magnetic bead hole on the sample plate, the nucleic acid in the biological sample to be extracted is decontaminated by the decontamination magnetic bead before being purified, and then the nucleic acid in the biological sample to be extracted is purified by the purification magnetic bead, so that the nucleic acid can be efficiently and high-quality extracted without centrifugal decontamination, and the operation is simple and the cost is low.

[0093] The above description has fully disclosed the specific embodiments of the utility model. It should be pointed out that any modification of the specific embodiments of the utility model by those skilled in the art does not deviate from the scope of the claims of the utility model. Accordingly, the scope of the claims of the utility model is not limited to the foregoing specific embodiments.

Claims

1. A magnetic bead extraction kit, characterized by, The application relates to a sample loading plate, which comprises at least one set of impurity removing magnetic bead holes, purification magnetic bead holes, storage holes and elution holes. The impurity removing magnetic bead holes comprise at least two reaction cavities completely separated by a separation film; the at least two reaction cavities comprise a magnetic bead solution cavity and a lysis solution cavity, the magnetic bead solution cavity contains an impurity removing magnetic bead solution, the impurity removing magnetic bead solution comprises impurity removing magnetic beads which are used for removing impurities in a nucleic acid in a biological sample to be extracted; the lysis solution cavity contains a lysis solution which is used for lysis of cell membranes to release nucleic acids; wherein the separation film is easy to break. The purification magnetic bead holes contain a purification magnetic bead solution, the purification magnetic bead solution comprises purification magnetic beads which are used for adsorbing nucleic acids in the biological sample to be extracted. The storage holes are used for temporarily storing the purification magnetic beads or the impurity removing magnetic beads in use. The elution holes contain an elution solution which is used for separating the nucleic acids from the purification magnetic beads. When the nucleic acids are extracted, the storage holes are used for temporarily storing the purification magnetic beads, and the purification magnetic bead holes are also used for temporarily storing the impurity removing magnetic beads after the impurities are adsorbed. Each set further comprises a cleaning hole used for storing a cleaning solution, or the storage holes further contain a cleaning solution.

2. The kit of claim 1, wherein The cleaning solution is used for cleaning the purification magnetic beads after the nucleic acids are adsorbed. The outer surface of the magnetic microsphere body of the impurity removing magnetic beads has at least one branched polymer with a functional group.

3. The kit according to claim 1 or 2, characterized in that, The depth of the impurity removing magnetic bead holes, the purification magnetic bead holes, the storage holes and the elution holes is 20-50 mm, and / or the cross section of each hole is square.

4. The kit of claim 1, wherein The sample loading plate sequentially distributes at least one set of the impurity removing magnetic bead holes, the purification magnetic bead holes, the storage holes and the elution holes along the row or column direction.

5. The kit of claim 1, wherein For one set of the impurity removing magnetic bead holes, the purification magnetic bead holes, the storage holes and the elution holes, the length of the impurity removing magnetic bead holes is at least twice the length of the purification magnetic bead holes, and the length of the purification magnetic bead holes is the same as the length of the storage holes and the elution holes.

6. The kit of claim 1, wherein The magnetic bead solution cavity of the impurity removing magnetic bead holes further contains a neutralizing solution; or the impurity removing magnetic bead holes further comprise a neutralizing solution cavity which is located between the magnetic bead solution cavity and the lysis solution cavity, wherein the neutralizing solution cavity contains a neutralizing solution.

7. The kit of claim 1, wherein The separation film horizontally separates the magnetic bead solution cavity and the lysis solution cavity, and the magnetic bead solution cavity is located below the lysis solution cavity.

8. The kit of claim 1, wherein The separation film is formed by pouring and solidifying a material with a solidification temperature higher than the ambient temperature after being heated to a molten state; and / or the material of the separation film is selected from paraffin.

9. The kit according to claim 7 or 8, characterized in that Further comprising a sealing film layer which is used for sealing the sample loading plate.

10. The kit of claim 1, wherein ​

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