CN107029452B - Method for removing plane rigid structure substance from sample - Google Patents
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- CN107029452B CN107029452B CN201710226145.7A CN201710226145A CN107029452B CN 107029452 B CN107029452 B CN 107029452B CN 201710226145 A CN201710226145 A CN 201710226145A CN 107029452 B CN107029452 B CN 107029452B
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/02—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor with moving adsorbents
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/42—Selective adsorption, e.g. chromatography characterised by the development mode, e.g. by displacement or by elution
- B01D15/424—Elution mode
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Abstract
The invention relates to a method for removing plane rigid structure substances from a sample, which comprises the steps of mixing or fully contacting a nucleic acid-solid phase material with the sample to be treated at normal temperature to combine plane rigid structure molecules in the sample to be treated with nucleic acid, and separating the sample to obtain the sample from which the plane rigid structure molecules are removed. The plane rigid structure molecules widely exist in living environments, such as benzopyrene in edible oil, aflatoxin in fresh milk and agricultural and sideline products, ethidium bromide in laboratory reagent, aristolochic acid in Chinese medicinal materials, and the like, and all have plane rigid structures. Such substances are often present in minute amounts, but present a greater health risk to humans and other animals. The invention is used for processing the sample containing the substances, thereby reducing the content of the plane rigid structure molecules in the sample to the minimum, having the advantages of high efficiency, convenient use, low cost and the like, and having wide application in the fields of food, medicine, sewage treatment and the like.
Description
Technical Field
The invention relates to a technical method for removing plane rigid structure molecules from a sample, which is suitable for the fields of food, medicines, sewage treatment and the like.
Background
Planar rigid molecules are used herein to refer to both molecules having a planar rigid configuration throughout the molecule and molecules having a planar rigid configuration in part of their structure. In these molecules, one or more aromatic ring systems are often present. These aromatic ring systems are bound in one plane by a conjugated system (e.g., pi-pi conjugation) to form a rigid planar structure.
This planar configuration, along with its hydrophobic nature, allows planar rigid molecules to interact with DNA double-stranded helices. It is believed that many planar rigid molecules are capable of intercalating between two adjacent pairs of base pairs of the double helix structure of DNA and also binding into the major and minor grooves of DNA. Such substances may have an effect on the function of DNA due to their ability to interact with DNA.
Ethidium bromide is a biochemical reagent frequently used in laboratories, has a plane rigid structure, is an ultra-strong DNA intercalator, and is a recognized DNA mutagen.
benzopyrene is believed to be a highly active indirect carcinogen that is capable of forming active derivatives with planar rigid structures in mammalian bodies under the action of enzymatic catabolism, after the benzopyrene enters the body, except for a small fraction of it being excreted in the body with feces, a fraction of it is converted to dozens of metabolites by mixed functional oxidase activation in liver and lung cell microsomes, a fraction of it is converted to epoxides, in particular to 7, 8-epoxides, which are metabolized to produce 7, 8-dihydrodihydroxy-9, 10-epoxybenzo [ a ] pyrene, which is believed to be a carcinogen, there are four isomers of this carcinogen, among which (+) -BP-7 β, 8 α -diol-9 α, 10 α -epoxide-benzo [ a ] pyrene, which has been shown to be most carcinogenic, forms a covalent bond with DNA, causing DNA damage, and if DNA cannot be repaired or repaired, cells may become cancerous.
Aflatoxin also has a planar rigid structure, is a toxic metabolite produced by aspergillus flavus, and is classified as a class 1 carcinogen by the cancer research institution of the World Health Organization (WHO). Aflatoxin is a substance with strong carcinogenicity, which is derived from various metabolites generated by hydroxylation in organisms. Aflatoxins are widely distributed in moldy grains and products thereof, particularly peanuts, peanut oil, corns and products thereof, milk and dairy products, and moldy feeds are found to contain more aflatoxins. Therefore, aflatoxins pose health risks to humans and animals.
Aristolochic acid is a component in Chinese medicinal materials, and Chinese pharmacopoeia and national drug standards are collected and used for preparing medicinal materials containing aristolochic acid, such as caulis Akebiae, radix Aristolochiae Fangchi, radix Aristolochiae, caulis Aristolochiae, herba Aristolochiae Mollissimae, radix Aristolochiae Mollissimae, and radix Aristolochiae Kaempferi. The plants have been widely used as crude drugs by processing and detoxifying traditional Chinese medicines, and plants containing aristolochic acid have been widely used internationally. Aristolochic acid I and its metabolite aristololactam have now been found to be nephrotoxic. The influence of aristolochic acid on the kidney is commonly known in the world, and is even called as Chinese herbal medicine nephropathy abroad, and Chinese scholars are called as aristolochic acid nephropathy. In addition, the metabolic process and metabolic enzyme research of aristolochic acid proves that aristolochic acid also has mutagenic and carcinogenic toxicity. Under the metabolic action, the produced intermediate aristolochia lactam nitrogen ions can be electrophilically combined with DNA base exocyclic amino to generate corresponding addition products, so that RAS genes and P53 genes are mutated to further induce tumors.
In addition, in natural drugs, coumarins also have a planar rigid structure. Although many coumarins are reported as effective physiologically active ingredients, many of these substances make it difficult to count the number of derivatives present in natural drug extracts, which can present health risks.
The removal of planar rigid molecules from samples prior to use or consumption is particularly important given that such molecules are present in large amounts in the living environment and pose health risks to mammals, including humans.
Due to their structural features, such molecules tend to associate with the DNA double helix structure. The inventor also proves through experiments that plane rigid molecules including benzopyrene and aflatoxin can be combined with DNA, so that the molecules can be removed by using the DNA.
In the application of DNA to a large volume of a sample containing planar rigid molecules, convenience of operation and economic cost need to be considered in addition to effectiveness. This would make the method more efficient and less costly if the DNA-planar rigid molecule complex could be easily separated and easily removed from the DNA molecule after the DNA has been applied to the sample to bind the planar rigid molecules therein, thereby allowing the DNA molecule to be reused.
Binding DNA to a solid phase would be a good strategy to achieve this goal. The attachment of DNA to a solid phase by covalent or non-covalent binding is a well established technique. The attachment of DNA to conventional solid phase materials such as cellulose, colloidal particles or agarose has been widely used. When DNA is applied to a sample in the form of a DNA-solid phase material, subsequent isolation becomes considerably easy, and the possibility of DNA remaining in the sample is also minimized. The DNA-solid phase material can be applied as an elution column through which the sample flows, wherein the planar rigid molecules are retained on the solid phase, allowing the sample to be purified. Alternatively, the mixture of DNA-solid phase material and sample can be centrifuged and/or filtered to separate the DNA-solid phase material along with the planar rigid molecules from the sample.
In addition to conventional solid phases such as cellulose, colloidal particles and agarose, another solid phase currently used for coupling to DNA is magnetic beads, such as streptavidin-coated magnetic beads, which are now widely used for biotinylation of nucleic acids. The superparamagnetic beads have streptavidin covalently attached to the surface, which can efficiently couple biotinylated DNA molecules. When DNA is applied in the form of DNA-magnetic beads, the DNA-magnetic beads can be easily separated from the sample together with the planar rigid molecules using magnetism.
After recovery of the DNA, the DNA may be denatured by raising the temperature. When the DNA is denatured, the double-stranded structure is opened, and the higher order structure of the DNA is destroyed, so that the binding force of the DNA molecule to the planar rigid molecule is weakened or completely lost, and thus the planar rigid molecule can be easily removed from the DNA molecule. After removal of the planar rigid molecules, the temperature can be lowered again to renature the DNA molecules and thus can be recycled for the next application. When applied in the form of DNA-solid phase materials such as DNA-magnetic beads, this subsequent DNA recycling operation will be facilitated.
When applying DNA-solid phase materials to remove planar rigid molecules, DNA may be subjected to more severe conditions, such as elevated temperature, friction, shear stress, etc. Such harsh conditions may cause premature denaturation of the DNA molecules, thereby reducing the binding force of DNA to the planar rigid molecules and thus reducing the removal efficiency. It is therefore advantageous for the DNA molecule to have a higher content of guanine (G) and uracil (C) bases. 3 hydrogen bonds can be formed between GC base pairs, so that the DNA molecules with higher GC base content can resist the harsh conditions in an enhanced way. Thus, in the examples of the present invention, the base content of the DNA molecule is high.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: the invention provides a method for removing a plane rigid structure substance from a sample, which can effectively reduce the content of plane rigid structure molecules in the sample and has the advantages of simple operation, low cost and high efficiency.
The technical scheme for solving the technical problems is as follows: a method of removing planar rigid structural material from a sample, comprising the steps of:
(1) mixing the nucleic acid-solid phase material with a sample to be treated thoroughly to obtain a mixture;
(2) separating the nucleic acid-solid phase material from the mixture to obtain a sample having the planar rigid structural molecules removed from the sample.
The sample to be treated is an oily solution or an aqueous solution.
The sample to be treated is an edible solution, an oil material, sewage or a medicinal extract.
Said solid phase material is cellulose, colloidal particles or agarose and wherein said separating comprises centrifugation and/or filtration and/or passing said sample to be treated through a column packed with said nucleic acid-solid phase material.
The solid phase material is magnetic beads, and wherein the separating comprises aspirating the nucleic acid-solid phase material with a magnetic device.
The nucleic acid is double-stranded DNA; the DNA sequence is GC-rich, i.e., the GC content of the DNA is 30% -100%.
The mixing method is stirring and mixing for 20 minutes to 5 hours, or the sample to be treated passes through a column filled with the nucleic acid-solid phase material.
The volume ratio of the nucleic acid-solid phase material to the sample to be treated is 1: 1-1: 500.
The method further comprises the step of recovering the nucleic acid-solid phase material for recycling; the specific steps for recovering the nucleic acid-solid phase material are as follows:
(1) increasing the temperature to denature the nucleic acid in the nucleic acid-solid phase material adsorbed with the planar rigid structure molecule;
(2) eluting or extracting the planar rigid structure molecules with a solvent;
(3) the temperature is lowered to renature the nucleic acid in the nucleic acid-solid phase material.
The specific steps for recovering the nucleic acid-solid phase material are as follows:
(1) raising the temperature of the nucleic acid-solid phase material adsorbed with the planar rigid structure molecules to 80-100 ℃, and slowly stirring for 20-100 minutes to denature the nucleic acid in the nucleic acid-solid phase material adsorbed with the planar rigid structure molecules;
(2) eluting or extracting planar rigid structure molecules in the nucleic acid-solid phase material by using a solvent; the solvent is hot water or an organic solvent with the temperature of 50-80 ℃;
(3) and reducing the temperature of the nucleic acid-solid phase material to room temperature to renature the nucleic acid in the nucleic acid-solid phase material to obtain the nucleic acid-solid phase material with the plane rigid structure molecules removed.
The invention mixes the DNA coupled with the solid phase with the sample to be processed at normal temperature, so that the plane rigid structure molecules in the sample are combined with the DNA. The solid phase-DNA-target complex is separated from the mixture by centrifugation (or using a magnetic stand if the solid phase is a magnetic bead) to remove the planar rigid structure molecules from the sample. Heating the separated solid phase-DNA-target compound to a DNA denaturation temperature to denature DNA, reducing the binding force between the DNA and the plane rigid structure molecules, and eluting the plane rigid molecules by using a solvent. Then the temperature is reduced to renature the DNA in the solid phase-DNA, thereby realizing the recycling of the solid phase-DNA and reducing the cost. The method is simple to operate, does not introduce new pollutants into the sample, can be recycled, and reduces the cost.
The plane rigid structure molecules widely exist in living environments, such as benzopyrene in edible oil, aflatoxin in fresh milk and agricultural and sideline products, ethidium bromide in laboratory reagent, aristolochic acid in Chinese medicinal materials, and the like, and all have plane rigid structures. Such substances are often present in minute amounts, but present a greater health risk to humans and other animals. The method of the invention can be used to treat samples containing such materials to minimize the amount of planar rigid structural molecules in the sample. The method has the advantages of high efficiency, convenient use, low cost and the like, and is widely applied to the fields of food, medicine, sewage treatment and the like.
Detailed Description
The invention mixes or fully contacts nucleic acid-solid phase material with sample to be processed at normal temperature, combines plane rigid structure molecule in the sample to be processed with nucleic acid, and separates out the sample, thereby obtaining the sample without plane rigid structure molecule, and the invention mainly has two technical proposals:
the first scheme is as follows:
(1) mixing the nucleic acid-solid phase material with a sample to be treated thoroughly to obtain a mixture; the mixing is to mix the nucleic acid-solid phase material and the sample to be treated for 20 minutes to 5 hours, so that the molecules with the plane rigid structure in the sample to be treated are combined with the nucleic acid; the volume ratio of the nucleic acid-solid phase material to the sample is 1: 1-1: 500;
(2) separating the nucleic acid-solid phase material from the mixture to obtain a sample having the planar rigid structural molecules removed from the sample; when the solid phase material is cellulose, colloidal particles or agarose, the separation is centrifugation and/or filtration; when the solid phase material is magnetic beads, the separation is performed by sucking out the nucleic acid-solid phase material by a magnetic device.
Scheme II:
(1) mixing the nucleic acid-solid phase material with a sample to be treated thoroughly to obtain a mixture; the mixing is that the sample to be processed passes through a column filled with the nucleic acid-solid phase material, so that the plane rigid structure molecules in the sample to be processed are combined with the nucleic acid; the height of the column is required to be not less than 40 cm.
(2) Separating the nucleic acid-solid phase material from the mixture, said separating also being a process of passing the sample to be treated through a column packed with the nucleic acid-solid phase material, to obtain a sample in which the planar rigid structural molecules are removed from the sample.
Example 1: and removing benzopyrene from the tea seed oil by using DNA-magnetic beads.
1. Weighing 100g of crude tea seed oil, detecting the content of benzopyrene to be 16.1 mu g/kg, and placing the sample in a 250ml glass container with a sealing cover.
2. 4-10ml of commercially available DNA-magnetic beads were added, and the cap was closed.
3. The mixture was stirred slowly at room temperature for 1 hour with a magnetic stirrer at a stirring speed of 50 rpm. During this period, the container is lifted by hand and shaken upside down several times at intervals of about 10 to 20 minutes to facilitate sufficient contact between the DNA-beads and the sample.
4. After 1 hour, the mixture was allowed to stand for about 1 minute to allow the DNA-beads to settle completely at the bottom of the vessel.
5. The sealing cover is opened, the magnetic frame is placed at the opening of the container, and most of DNA-magnetic beads in the container are adsorbed on the magnetic frame. The reaction was kept for 5 minutes and the oil droplets remaining on the DNA-beads were allowed to drain.
6. The magnetic holder is removed from the container opening and the DNA-beads on the magnetic holder are removed. The magnetic frame is again placed on the container opening, and the DNA-magnetic beads in the container are completely aspirated away.
7. And detecting the benzopyrene content of the treated tea seed oil sample again, wherein the detection value is 5.4 mug/kg.
The tea seed oil selected in this example had a benzopyrene content of 16.1. mu.g/kg, and the oil treated with a common activated carbon adsorption method had a benzopyrene content of about 10. mu.g/kg. However, the amount of oil and fat loss is 1 to 2 times of the amount of the activated carbon, the activated carbon after adsorbing benzopyrene is difficult to treat, and the benzopyrene can be leaked into the air again due to analysis or calcination. The method for removing benzopyrene by using immobilized DNA is used for treating under the conditions, the amount of the benzopyrene after removal can be reduced to 5.4 mug/kg, and meanwhile, the subsequent operation is simple without any residue.
Therefore, the method for removing benzopyrene by using immobilized DNA has the advantages of good removal effect, obvious reduction of grease loss, simple operation and low cost.
Of course, it is obvious that the solid phase in the DNA-magnetic beads in this embodiment can be changed to other common solid phases, and similarly good effects can be obtained.
Example 2: the ethidium bromide is removed from the aqueous solution using DNA-magnetic beads.
China has about 2000 colleges and universities, and 10 thousands of chemical laboratories exist in various chemical laboratories, and a large number of teachers and students perform various chemical experiments every day to generate a large amount of waste liquid, wherein ethidium bromide is one of the most harmful pollutants. In this example, a fixed amount of ethidium bromide was added to a volume of water as a sample of the contaminated water, and the ethidium bromide was removed using the method of the present invention to demonstrate the utility of the present invention in wastewater treatment. The embodiment comprises the following steps:
1. ethidium Bromide (EB) was added to 500ml of purified water in a glass container with a sealing lid to prepare a wastewater sample having an ethidium bromide content of 50 mg/l.
2. 4-10ml of commercially available DNA-magnetic beads were added, and the cap was closed.
3. The mixture was stirred slowly with a magnetic stirrer at a stirring speed of 50 rpm for about 20 minutes at room temperature.
4. After 20 minutes, the mixture was allowed to stand for about 1 minute to allow the DNA-beads to settle completely at the bottom of the vessel.
5. And opening the sealing cover, and placing the container on the opening of the container by using a magnetic frame to ensure that all the DNA-magnetic beads in the container are adsorbed on the magnetic frame. The reaction mixture was kept for 5 minutes, and the residual water on the DNA-beads was drained.
6. The magnetic holder is removed from the container opening and the DNA-beads on the magnetic holder are removed.
7. And detecting the content of the ethidium bromide in the treated water by utilizing the characteristic absorption peak of the ethidium bromide at 480 nm. The ethidium bromide content of the sample was found to be undetectable.
In this example, because ethidium bromide has a very strong affinity for DNA, all of the ethidium bromide in the sample can be adsorbed with relatively little DNA. In addition, since the DNA-magnetic beads are completely removed from the sample, it can be used to completely remove ethidium bromide from a contaminated aqueous sample.
It will be apparent, of course, that the present invention can be used to treat not only water bodies contaminated with ethidium bromide, but also water bodies contaminated with other planar rigid molecular species. According to the affinity degree of the pollution molecules and the DNA, the dosage of the DNA-magnetic beads can be correspondingly adjusted, and the good effect can be achieved. In addition, the solid phase material used in this embodiment is not limited to magnetic beads, and other solid phases commonly used for DNA immobilization are also applicable to the present invention.
The present invention can ensure that an effective amount of the planar rigid structural molecules in the nucleic acid-solid phase material, which depends on the length and GC content of the nucleic acid sequence in the nucleic acid-solid phase material and the nature of the planar rigid structural molecules, are bound to DNA by defining the volume ratio of the nucleic acid-solid phase material to the sample. It is generally required that the volume ratio (V/V) of the nucleic acid-solid phase material to the sample is 1:1 to 1:500, and may be 1:1, 1:5, 1:10, 1:20, 1:30, 1:40, 1:50, 1:100, 1:150, 1:200, 1:250, 1:300, 1:350, 1:400 or 1: 500.
Example 3: aflatoxin B1 was removed from the solution using a DNA-dextran bead column.
Because crops are often infected with aspergillus flavus, the agricultural byproducts possibly contain aflatoxin B1, which is a very toxic pollutant. This example shows the method of the present invention to remove aflatoxin B1 from a sample. The embodiment comprises the following steps:
1. a commercially available aflatoxin B1 methanol standard was formulated into 5. mu.g/ml samples in 30ml methanol/water (4/1, v/v) solution.
2. Commercially available DNA-dextran microbeads were packed in a column with a column height of 50cm and equilibrated at room temperature for 30 minutes.
3. The column was washed 2 times with 10ml portions of deionized water and the liquid was allowed to flow out at a flow rate of 1-5 drops/sec.
4. And adding the sample into a purification column, and regulating the flow to 1-6 drops/second until the sample completely flows out of the purification column to obtain the sample from which the planar rigid structure molecules are removed.
5. The column was washed 3 times with 2-10 ml of distilled water each time.
6. 20ml of methanol was added and the eluted product was collected.
7. And detecting the content of aflatoxin B1 in the eluted product.
The purification column was determined to adsorb 65.6% of aflatoxin B1 in the sample. Although the Aspergillus flavus removal efficiency still needs to be improved, in view of the simple operation of the method of the present invention, the DNA-solid phase material can be recycled after purification recovery, and the sample purified by the column is loaded again, and the above steps 4 to 7 are repeated. The aflatoxin content in the sample was determined to be further reduced to 21.1%. Therefore, by treating the sample containing aflatoxin by the method of the invention for multiple times, the toxin content can be greatly reduced.
Of course, it is obvious that the solid phase in the DNA-dextran bead in this embodiment can be changed to other common solid phases, such as magnetic beads, and similarly good effect can be obtained.
The DNA in each example of the present invention is a double-stranded DNA; the sequence of the DNA is GC-rich and may be 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99%, or even 100% GC content, i.e., 30% -100% GC content in the DNA.
Example 4: recycling of DNA-magnetic beads adsorbed with planar rigid molecular materials
The invention is characterized in that the DNA-solid phase material can be recycled to reduce the cost. The following examples will demonstrate the recycling process of the present invention. The method comprises the following specific steps:
1. 2ml of the DNA-magnetic beads adsorbed with Ethidium Bromide (EB) (EB content 6.3 mg) from example 2 were collected and placed in a 500ml glass flask in purified water.
2. The temperature in the glass flask was raised to 90 degrees celsius with a water bath and stirred slowly for 30 minutes.
3. The DNA-beads were aspirated from the glass flask by a magnetic holder and rapidly washed with hot water for 30 seconds.
4. The DNA-magnetic beads were removed from the magnetic stand, stored at room temperature for about 30 minutes, and then assayed for ethidium bromide content.
The detection result shows that ethidium bromide in the DNA-magnetic beads can not be detected, so that the ethidium bromide can be completely removed from the DNA-magnetic beads by eluting the ethidium bromide with an eluent at the denaturation temperature of the DNA, and the DNA-magnetic beads can be recycled.
In step 3 of example 4 of the present invention, in addition to hot water elution of the planar rigid molecules, organic solvents such as methanol, ethanol, acetone, chloroform, etc. may be used.
Claims (3)
1. A method of removing planar rigid structural material from a sample, comprising: the method comprises the following steps:
(1) mixing the nucleic acid-solid phase material with a sample to be treated thoroughly to obtain a mixture; the mixing is to mix the nucleic acid-solid phase material and the sample to be treated for 20 minutes to 5 hours, so that the molecules with the plane rigid structure in the sample to be treated are combined with the nucleic acid; the volume ratio of the nucleic acid-solid phase material to the sample is 1: 1-1: 500; the sample to be processed is tea seed oil containing a planar rigid structure molecule benzopyrene or aqueous solution containing a planar rigid structure molecule ethidium bromide; the nucleic acid-solid phase material is DNA-magnetic beads;
(2) separating the nucleic acid-solid phase material from the mixture, i.e. aspirating the nucleic acid-solid phase material with a magnetic device, to obtain a sample in which the planar rigid structural molecules are removed from the sample;
the method further comprises the step of recovering the nucleic acid-solid phase material for recycling; the specific steps for recovering the nucleic acid-solid phase material are as follows:
(1) increasing the temperature to denature the nucleic acid in the nucleic acid-solid phase material adsorbed with the planar rigid structure molecule;
(2) eluting or extracting the planar rigid structure molecules with a solvent;
(3) the temperature is lowered to renature the nucleic acid in the nucleic acid-solid phase material.
2. A method of removing planar rigid structural material from a sample according to claim 1, wherein: the nucleic acid is double-stranded DNA; the DNA sequence is GC-rich, i.e., the GC content of the DNA is 30% -100%.
3. A method of removing planar rigid structural material from a sample according to claim 1, wherein: the specific steps for recovering the nucleic acid-solid phase material are as follows:
(1) raising the temperature of the nucleic acid-solid phase material adsorbed with the planar rigid structure molecules to 80-100 ℃, and slowly stirring for 20-100 minutes to denature the nucleic acid in the nucleic acid-solid phase material adsorbed with the planar rigid structure molecules;
(2) eluting or extracting planar rigid structure molecules in the nucleic acid-solid phase material by using a solvent; the solvent is hot water or an organic solvent with the temperature of 50-80 ℃;
(3) and reducing the temperature of the nucleic acid-solid phase material to room temperature to renature the nucleic acid in the nucleic acid-solid phase material to obtain the nucleic acid-solid phase material with the plane rigid structure molecules removed.
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| CN103657145A (en) * | 2008-08-01 | 2014-03-26 | 生物风险公司 | Devices and methods for the purification, isolation, desalting or buffer/solvent exchange of substances |
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| US20020090628A1 (en) * | 1998-11-23 | 2002-07-11 | Gray Joe W. | Y chromosome specific nucleic acid probe and method for determining the Y chromosome in situ |
| CN101013111B (en) * | 2007-02-13 | 2010-05-19 | 段为钢 | Chart sifting method of effective composition in traditional Chinese medicine |
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| JP2004351336A (en) * | 2003-05-29 | 2004-12-16 | Canon Inc | Dna hybrid and environment cleaning system using the same |
| CN1968745A (en) * | 2004-06-15 | 2007-05-23 | 佳能株式会社 | Structure designed for adsorption of DNA intercalators |
| CN103657145A (en) * | 2008-08-01 | 2014-03-26 | 生物风险公司 | Devices and methods for the purification, isolation, desalting or buffer/solvent exchange of substances |
| CN103087829A (en) * | 2013-02-25 | 2013-05-08 | 厦门大学 | Method for removing furocoumarin from essential oil |
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