Application of novel adsorption material to extraction of nucleic acid

Abstract

The invention discloses application of a novel adsorption material to extraction of nucleic acid, the adsorption material is macroporous adsorption resin, and an application method for extraction of nucleic acid comprises the following steps: S1, collecting a biological sample or an environmental sample, adding a lysis solution, and releasing nucleic acid including DNA, RNA and plasmids; s2, adding macroporous adsorption resin for adsorption, fully oscillating, centrifuging to remove the resin and adsorbate thereof, and retaining liquid; s3, macroporous adsorption resin is added for adsorption, full oscillation is performed, the resin and adsorbate thereof are removed through centrifugation, and liquid is reserved.

Description

Technical Field

[0001] The present invention relates to the technical field of application of a novel adsorption material for extracting nucleic acid, and in particular to the application of a novel adsorption material for extracting nucleic acid. Background Art

[0002] Currently, the silica-based membrane (spin column) method relies on a high-salt buffer to adsorb nucleic acids to the silica membrane. This method is cumbersome, has high reagent costs, and cannot achieve high-throughput extraction. The magnetic bead method uses surface-modified magnetic beads to bind nucleic acids and relies on magnetic field separation, requiring sophisticated equipment and high bead costs. It also relies on chemical reagents (such as isopropanol), has multiple steps, is time-consuming, can lead to significant nucleic acid loss, and has low extraction efficiency. Some materials are also incompatible with complex samples (such as blood and soil). Summary of the Invention

[0003] In order to solve one or some technical problems existing in the prior art, the purpose of this application is to provide an application of a new adsorption material for extracting nucleic acids, which simplifies the operation process of traditional extraction methods and shortens the nucleic acid extraction time.

[0004] In order to solve the above existing technical problems, one of the objectives of this application is achieved by adopting the following technical solutions: A novel adsorption material is used to extract nucleic acids. The adsorption material is a macroporous adsorption resin, and the method for extracting nucleic acids is as follows: S1, collect biological samples or environmental samples, add lysis buffer, and release nucleic acids including DNA, RNA, and plasmids; S2, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S3, add macroporous adsorption resin for adsorption, shake thoroughly, centrifuge to remove the resin and its adsorbate, and retain the liquid.

[0005] Furthermore, when the sample is any one of animals, plants, microorganisms, soil, and feces, the steps are as follows: S1: Collect animal, plant, microbial, soil, feces, water or environmental samples, add corresponding lysis buffer to release or disperse the DNA in the samples; S2, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S3, the resulting liquid is the DNA sample.

[0006] Furthermore, when the sample is any one of animal tissue, blood or bacterial sample, the steps are as follows: S1, collect animal tissue, blood or bacterial samples, add guanidine isothiocyanate lysis buffer to release RNA; S2, add phenol-chloroform solution, mix well, and centrifuge to obtain the supernatant; S3, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S4, the resulting liquid is the RNA sample.

[0007] Furthermore, when the sample is any one of plant tissue and fungal sample, the steps are as follows: S1: Collect plant tissue and fungal samples, add liquid nitrogen and grind them into powder. Then, add guanidine isothiocyanate lysis solution to the powder to release RNA. S2, add phenol-chloroform solution, mix well, and centrifuge to obtain the supernatant; S3, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S4, the resulting liquid is the RNA sample.

[0008] Furthermore, when the sample is an Escherichia coli sample containing a plasmid, the steps are as follows: S1, collect the E. coli sample containing the plasmid, add diluent, and disperse the microorganisms; S2, add SDS / sodium hydroxide and potassium acetate to release plasmid DNA, mix well and centrifuge to obtain the supernatant; S3, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S4, the obtained liquid is the plasmid DNA sample.

[0009] Furthermore, when the sample is a PCR product sample, the steps are as follows: S1, collect PCR product samples; S2, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S3, the resulting liquid is the nucleic acid sample.

[0010] Furthermore, when the sample is a DNA / RNA sample containing various impurities, the steps are as follows: S1, collect various DNA / RNA samples containing impurities and add diluent to dissolve or dilute the samples; S2, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S3, the resulting liquid is a pure nucleic acid sample.

[0011] Compared with the prior art, the present invention has the following beneficial effects: (1) The method of this application uses macroporous adsorption resin to adsorb various impurities in the sample, and then separates the resin bound to the impurities from the sample by centrifugation. The resulting clear solution is the nucleic acid sample, which can be directly used in downstream experiments. This simplifies the operation process of traditional extraction methods, shortens the nucleic acid extraction time, increases the total amount and purity of nucleic acid extracted, and is beneficial for downstream applications. DETAILED DESCRIPTION

[0012] Below, the present application is further described in conjunction with specific implementation methods. It should be noted that, under the premise of no conflict, the various embodiments or technical features described below can be arbitrarily combined to form new embodiments.

[0013] In the description of this application, it should be understood that the terms "up", "down", "left", "right", etc., indicating directions or positional relationships, are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific direction, be constructed and operated in a specific direction. Therefore, it should not be understood as a limitation on this application.

[0014] The terms "first", "second", etc. in this application are used to distinguish similar objects, and are not used to describe a specific order or sequence. It should be understood that the data used in this way can be interchangeable under appropriate circumstances, so that the embodiments of the present application can be implemented in an order other than those described here, and the objects distinguished by "first", "second", etc. are generally of one type, and the number of objects is not limited. For example, the first object can be one or more. In addition, "and / or" in the specification and claims represents at least one of the connected objects, and the character " / " generally indicates that the previously associated objects are an "or" relationship of application of a new adsorption material for extracting nucleic acids. Embodiment 1:

[0015] A novel adsorption material is used to extract nucleic acids. The adsorption material is a macroporous adsorption resin, and the method for extracting nucleic acids is as follows: S1, collect biological samples or environmental samples, add lysis buffer, and release nucleic acids including DNA, RNA, and plasmids; S2, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S3, add macroporous adsorption resin for adsorption, shake thoroughly, centrifuge to remove the resin and its adsorbate, and retain the liquid. The method of the present application uses macroporous adsorption resin to adsorb various impurities in the sample, and separates the resin bound to the impurities from the sample by centrifugation. The resulting clear solution is a nucleic acid sample and can be directly used in downstream experiments. It simplifies the operating procedures of traditional extraction methods, shortens the nucleic acid extraction time, increases the total amount and purity of nucleic acid extraction, is beneficial to downstream applications, and has high compatibility.

[0016] Furthermore, when the sample is any of animals, plants, microorganisms, soil, and feces, the steps are as follows: S1: Collect animal, plant, microbial, soil, feces, water or environmental samples, add corresponding lysis buffer to release or disperse the DNA in the samples; S2, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S3, the resulting liquid is the DNA sample.

[0017] Furthermore, when the sample is any of animal tissue, blood or bacterial sample, the steps are as follows: S1, collect animal tissue, blood or bacterial samples, add guanidine isothiocyanate lysis buffer to release RNA; S2, add phenol-chloroform solution, mix well, and centrifuge to obtain the supernatant; S3, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S4, the resulting liquid is the RNA sample.

[0018] Furthermore, when the sample is any one of plant tissue and fungal sample, the steps are as follows: S1: Collect plant tissue and fungal samples, add liquid nitrogen and grind them into powder. Then, add guanidine isothiocyanate lysis solution to the powder to release RNA. S2, add phenol-chloroform solution, mix well, and centrifuge to obtain the supernatant; S3, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S4, the resulting liquid is the RNA sample.

[0019] Furthermore, when the sample is an E. coli sample containing a plasmid, the steps are as follows: S1, collect the E. coli sample containing the plasmid, add diluent, and disperse the microorganisms; S2, add SDS / sodium hydroxide and potassium acetate to release plasmid DNA, mix well and centrifuge to obtain the supernatant; S3, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S4, the obtained liquid is the plasmid DNA sample.

[0020] Furthermore, when the sample is a PCR product sample, the steps are as follows: S1, collect PCR product samples; S2, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S3, the resulting liquid is the nucleic acid sample.

[0021] Furthermore, when the sample is a DNA / RNA sample containing various impurities, the steps are as follows: S1, collect various DNA / RNA samples containing impurities and add diluent to dissolve or dilute the samples; S2, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S3, the resulting liquid is a pure nucleic acid sample.

[0022] The above-mentioned embodiments are only preferred embodiments of the present application and cannot be used to limit the scope of protection of the present application. Any non-substantial changes and replacements made by technicians in this field based on the present application shall fall within the scope of protection required by the present application.

Claims

1. Application of a novel adsorption material for extracting nucleic acid, characterized in that: The adsorption material is a macroporous adsorption resin, and the application method for extracting nucleic acid is as follows: S1, collect biological samples or environmental samples, add lysis buffer, and release nucleic acids including DNA, RNA, and plasmids; S2, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S3, add macroporous adsorption resin for adsorption, shake thoroughly, centrifuge to remove the resin and its adsorbate, and retain the liquid.

2. The use of a novel adsorption material for extracting nucleic acid according to claim 1, characterized in that: When the sample is any of animals, plants, microorganisms, soil, or feces, the steps are as follows: S1: Collect animal, plant, microbial, soil, feces, water or environmental samples, add corresponding lysis buffer to release or disperse the DNA in the samples; S2, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S3, the resulting liquid is the DNA sample.

3. The use of a novel adsorption material for extracting nucleic acid according to claim 2, characterized in that: When the sample is any of animal tissue, blood, or bacterial samples, the steps are as follows: S1, collect animal tissue, blood or bacterial samples, add guanidine isothiocyanate lysis buffer to release RNA; S2, add phenol-chloroform solution, mix well, and centrifuge to obtain the supernatant; S3, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S4, the resulting liquid is the RNA sample.

4. The use of a novel adsorption material for extracting nucleic acid according to claim 3, characterized in that: When the sample is a plant tissue or fungus sample, the steps are as follows: S1: Collect plant tissue and fungal samples, add liquid nitrogen and grind them into powder. Then, add guanidine isothiocyanate lysis solution to the powder to release RNA. S2, add phenol-chloroform solution, mix well, and centrifuge to obtain the supernatant; S3, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S4, the resulting liquid is the RNA sample.

5. The use of a novel adsorption material for extracting nucleic acid according to claim 4, characterized in that: When the sample is an E. coli sample containing a plasmid, the steps are as follows: S1, collect the E. coli sample containing the plasmid, add diluent, and disperse the microorganisms; S2, add SDS / sodium hydroxide and potassium acetate to release plasmid DNA, mix well and centrifuge to obtain the supernatant; S3, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S4, the obtained liquid is the plasmid DNA sample.

6. The use of a novel adsorption material for extracting nucleic acid according to claim 5, characterized in that: When the sample is a PCR product sample, the steps are as follows: S1, collect PCR product samples; S2, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S3, the resulting liquid is the nucleic acid sample.

7. The use of a novel adsorption material for extracting nucleic acid according to claim 6, characterized in that: When the sample is a DNA / RNA sample containing various impurities, the steps are as follows: S1, collect various DNA / RNA samples containing impurities and add diluent to dissolve or dilute the samples; S2, adding macroporous adsorption resin for adsorption, shaking thoroughly, centrifuging to remove the resin and its adsorbate, and retaining the liquid; S3, the resulting liquid is a pure nucleic acid sample.