CN114507661A - Rapid extraction method of genomic DNA of acer negundo - Google Patents
Rapid extraction method of genomic DNA of acer negundo Download PDFInfo
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- 238000000605 extraction Methods 0.000 title claims abstract description 17
- 244000046151 Acer negundo Species 0.000 title description 3
- 235000004422 Acer negundo Nutrition 0.000 title description 3
- 239000002244 precipitate Substances 0.000 claims abstract description 40
- 241000208140 Acer Species 0.000 claims abstract description 32
- 238000000034 method Methods 0.000 claims abstract description 30
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 28
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 25
- 239000006228 supernatant Substances 0.000 claims abstract description 21
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 claims abstract description 19
- 238000002156 mixing Methods 0.000 claims abstract description 16
- 239000000243 solution Substances 0.000 claims abstract description 16
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims abstract description 14
- PHTQWCKDNZKARW-UHFFFAOYSA-N isoamylol Chemical compound CC(C)CCO PHTQWCKDNZKARW-UHFFFAOYSA-N 0.000 claims abstract description 12
- 241000219240 Acer pictum subsp. mono Species 0.000 claims abstract description 11
- 239000011259 mixed solution Substances 0.000 claims abstract description 11
- 239000000843 powder Substances 0.000 claims abstract description 11
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000011536 extraction buffer Substances 0.000 claims abstract description 6
- 238000005406 washing Methods 0.000 claims abstract description 5
- 241000510091 Quadrula quadrula Species 0.000 claims abstract description 4
- 238000007605 air drying Methods 0.000 claims abstract description 4
- 230000008014 freezing Effects 0.000 claims abstract description 4
- 238000007710 freezing Methods 0.000 claims abstract description 4
- 238000000227 grinding Methods 0.000 claims abstract description 4
- 241000931515 Acer palmatum Species 0.000 claims description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 8
- 239000008346 aqueous phase Substances 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 7
- 229910000831 Steel Inorganic materials 0.000 claims description 6
- 239000010959 steel Substances 0.000 claims description 6
- 239000008367 deionised water Substances 0.000 claims description 5
- 229910021641 deionized water Inorganic materials 0.000 claims description 5
- 239000012071 phase Substances 0.000 claims description 5
- 229910052757 nitrogen Inorganic materials 0.000 claims description 4
- 238000005336 cracking Methods 0.000 claims description 3
- 239000003517 fume Substances 0.000 claims description 3
- 230000007306 turnover Effects 0.000 claims 1
- 238000007400 DNA extraction Methods 0.000 abstract description 4
- 108091092562 ribozyme Proteins 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 150000004676 glycans Chemical class 0.000 description 4
- 229920001282 polysaccharide Polymers 0.000 description 4
- 239000005017 polysaccharide Substances 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 238000000246 agarose gel electrophoresis Methods 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 150000002989 phenols Chemical class 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 241001143500 Aceraceae Species 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 239000003147 molecular marker Substances 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- 238000009394 selective breeding Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 241000694401 Acer maximowiczianum Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
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- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
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- 230000008569 process Effects 0.000 description 1
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- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- SWGJCIMEBVHMTA-UHFFFAOYSA-K trisodium;6-oxido-4-sulfo-5-[(4-sulfonatonaphthalen-1-yl)diazenyl]naphthalene-2-sulfonate Chemical compound [Na+].[Na+].[Na+].C1=CC=C2C(N=NC3=C4C(=CC(=CC4=CC=C3O)S([O-])(=O)=O)S([O-])(=O)=O)=CC=C(S([O-])(=O)=O)C2=C1 SWGJCIMEBVHMTA-UHFFFAOYSA-K 0.000 description 1
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- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
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Abstract
The invention discloses a rapid extraction method of genomic DNA of acer mono, which comprises the following steps: preparing 1-2 g of northeast maple green leaves, quickly freezing, and grinding maple leaves into powder by a high-throughput tissue grinder; adding the prepared mixed solution of PVP and beta-mercaptoethanol into a centrifugal tube containing powder, uniformly mixing by shaking, centrifuging after ice bath, and removing supernatant to obtain precipitate; adding prepared CTAB extraction buffer solution containing RNA enzyme into the precipitate, and centrifuging to obtain supernatant; adding chloroform into the supernatant, shaking, mixing uniformly, and centrifuging to obtain an upper-layer water phase; adding isoamyl alcohol into the upper water phase, uniformly mixing and centrifuging to obtain a precipitate; washing the obtained precipitate twice with 70% ethanol, and air drying the ethanol to obtain the genomic DNA of the northeast maple leaf blade. The method has the advantages of high DNA extraction quality, low cost and short time consumption, and remarkably improves the extraction efficiency and the DNA purity of the northeast maple DNA.
Description
Technical Field
The invention belongs to the technical field of molecular biology, and particularly relates to a rapid extraction method of genomic DNA of acer negundo.
Background
Northeast maple also called Acer nikoense, Acer guayense, belonging to Acer genus plant of Aceraceae family, tall deciduous tree; the bark is gray to grey brown, coarse and has fine lines and longitudinal cracks; three-out compound leaves are opposite; the autumn leaves are pink or purple red, so the method has great ornamental value, is an important leaf-viewing tree species in the northeast China autumn, and is also an ideal tree species for the construction of urban garden landscapes. However, the population structure is unstable, natural propagation is difficult, the population quantity has fluctuation, meanwhile, the number of protected individuals is small, the influence of artificial activities and plant diseases and insect pests is large, natural updating is difficult, and the survival and protection conditions are not optimistic; at present, wild planting resources are increasingly reduced, and when northeast maples are transplanted into urban gardens, leaf color change often cannot be stably expressed compared with wild states, and the problems of short leaf-viewing period, non-uniform leaf color change period and the like often occur. Therefore, researches on genetic diversity of the germplasm resources of the acer northeast are increasingly emphasized by researchers at the DNA molecular level, and the genetic improvement process of the acer northeast can be effectively accelerated by adopting a conventional breeding method and a molecular marker-assisted selective breeding method.
As is well known, the extraction of genomic DNA is the very critical 'first step' for molecular marker-assisted selective breeding, and the rapid and efficient extraction of target DNA saves more effective and precious time for subsequent molecular tests. At present, DNA extraction methods are various, and a kit method, a CTAB method, an SDS method and the like are more mainstream, but the advantages and disadvantages of various methods are more obvious, the cost is high, the steps are more, the operation is complicated, the extraction time is too long, toxic reagents are probably generated, and the like, and the defects are more prominent in different extraction methods.
In actual experiments, the acer mono leaf tissue cells contain more substances such as polysaccharides, phenols, proteins and the like. In the initial stage of the test, when the traditional CTAB method is adopted to extract the tissue genome DNA of the acer mono leaf, the extracted DNA has extremely poor quality and is extremely viscous due to more impurities, and the gun head is blocked, so that the subsequent test can hardly be met. The subsequent reagent kit method is adopted to extract the genomic DNA of the acer northeast maple, but the concentration of the extracted DNA is low, the price of the reagent kit is high, and the test cost is high. Therefore, it is important to improve the existing extraction method to obtain a DNA extraction method with low cost, time and labor saving and high quality.
Disclosure of Invention
In order to overcome the problems, the invention provides a rapid extraction method of genomic DNA of acer northeast, which is improved on the basis of the traditional CTAB method and realizes the breakthrough of extraction of the genomic DNA of the acer northeast with high quality, high concentration, low cost, easy operation and short time consumption.
The technical scheme adopted by the invention is as follows:
a rapid extraction method of genomic DNA of acer northeast comprises the following steps:
the method comprises the following steps: putting a sterilized steel ball into a sterilized 2ml centrifuge tube, cutting the acer palmatum leaves into pieces, adding the cut acer palmatum leaves into the centrifuge tube, putting the sterilized steel ball into the centrifuge tube, quickly putting the acer palmatum leaves into liquid nitrogen for freezing, and grinding the acer palmatum leaves into powder by a high-throughput tissue grinder;
step two: checking whether the centrifugal tube has cracks or not, and transferring the northeast maple leaf powder into a new tube if the centrifugal tube has cracks; adding a mixed solution of 2% (g/ml) soluble PVP and 1% (mu L/ml) beta-mercaptoethanol with the same volume into the powder, shaking and uniformly mixing, carrying out ice bath for 10min, centrifuging, and then removing supernate to obtain a precipitate;
step three: discarding the supernatant, leaving a precipitate, adding a prepared CTAB extraction buffer solution containing RNaseA into the precipitate, shaking and uniformly mixing, putting into a 65 ℃ water bath kettle, carrying out water bath cracking for 10min, and centrifuging to obtain a supernatant;
step four: the supernatant is left, the precipitate is discarded, chloroform is added into the supernatant, and the supernatant is centrifuged after shaking and mixing to obtain an upper aqueous phase;
step five: keeping the upper aqueous phase, discarding the lower aqueous phase, adding isoamylol into the upper aqueous phase, uniformly mixing and centrifuging to obtain a precipitate;
step six: discarding the supernatant, leaving a precipitate, washing the precipitate with 70% ethanol twice, drying the ethanol in the air, and dissolving the precipitate with deionized water to obtain the genomic DNA solution of the acer negundo.
Wherein the northeast maple leaves selected in the step one are healthy green leaves, the petioles are removed when the leaves are cut into pieces, the weight is 1g-2g, and the high-throughput tissue grinder is set to grind for 2min at 65 Hz.
Wherein the volume of the mixed solution of 2 percent (g/ml) of soluble PVP and 1 percent (mu L/ml) of beta-mercaptoethanol added in the step two is 700 mu L (the mixed solution is placed in a refrigerator at 4 ℃ in a dark place) and is placed on ice in an ice bath, and the mixed solution is centrifuged at 12500rpm for 3 min.
Wherein, CTAB extraction buffer solution in the third step is 700 μ L of CTAB solution which is preheated (the CTAB extraction buffer solution can generate precipitation due to too low temperature), and 10 μ L of 10mg/ml RNaseA is added into 1ml of CTAB; turning upside down and mixing once every 5min while water bath; centrifuging at 12500rpm for 5min at 15 deg.C.
And the fourth step is to transfer the supernatant into a new sterile 1.5ml centrifuge tube, add 600. mu.L chloroform into the centrifuge tube, shake for 30s, and centrifuge for 5min at 12500 rpm.
And step five, specifically, taking 400 mu L of the upper-layer water phase to a new sterile 1.5ml centrifugal tube, adding 800 mu L of isoamyl alcohol into the centrifugal tube, slightly turning the centrifugal tube upside down, and centrifuging the centrifugal tube at 13500rpm for 15min after flocculent precipitates appear.
Wherein the sixth step is specifically that 700 microliter of 70% ethanol is added into the centrifuge tube containing the precipitate, the precipitate is washed by turning over the centrifuge tube for 30s, and the step is repeated twice; pouring out excessive ethanol in the tube, keeping precipitate, centrifuging at 12500rpm for 2min, sucking out excessive liquid with a gun head, and air drying in a fume hood; dissolving the dried precipitate with 50 μ L sterilized deionized water to obtain genomic DNA solution of Acer mono Maxim.
When the traditional CTAB method is used for extracting genomic DNA of the acer northeast, the extracted DNA is difficult to remove because the contents of phenols, pigments, proteins, polysaccharides and other substances in the leaf tissues of the acer northeast are too high, the extracted DNA is extremely viscous, and an electrophoresis strip is seriously dragged. Therefore, compared with the prior art, the invention has the beneficial effects that: compared with the classical CTAB method, the rapid and efficient extraction method of the genomic DNA of the acer northeast provided by the invention has the advantages that 2% (g/ml) of soluble PVP and 1% (mu L/ml) of beta-mercaptoethanol mixed solution with the same volume are added in the first step, polyphenol and polysaccharide in a sample can be effectively removed, then the prepared CTAB solution is further used for carrying out secondary removal on substances such as protein, phenols and polysaccharide in the sample, then chloroform is used for further removing protein pollution, isoamyl alcohol is used for precipitating DNA, and then 70% alcohol is used for washing and precipitating.
Drawings
FIG. 1 is the agarose gel electrophoresis test chart of DNA of 9 acer northeast maple samples extracted according to the method of example 1;
FIG. 2 is a drawing showing DNA agarose gel electrophoresis detection of 6 samples of acer northeast maple extracted according to the conventional CTAB method.
Detailed Description
In order to facilitate understanding of the present invention, the present invention will be described more fully and in detail with reference to the following specific examples, but the scope of the present invention is not limited to the specific examples.
Unless otherwise defined, all terms of art used hereinafter are consistent with their commonly understood meaning to those skilled in the art. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention.
Unless otherwise specifically stated, various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or can be prepared by existing methods.
Example 1
A rapid extraction method of genomic DNA of acer northeast comprises the following steps:
the method comprises the following steps: picking fresh and healthy green leaves of the acer mono in the field, storing the leaves with an ice box, and immediately transferring the leaves to the temperature of minus 30 ℃ for storage for later use; when the experiment is carried out, 1.5g of green leaves are quickly taken, cut into pieces and then added into a sterilized 2ml centrifuge tube (one sterilized steel ball is firstly added into the tube, and one steel ball is then added after the crushed leaves are added), then the tube is quickly put into liquid nitrogen for freezing, and then the liquid nitrogen is used for grinding for 2min by a high-flux tissue grinder with the frequency of 65Hz to form powder;
step two: checking whether the centrifugal tube has cracks or not, and transferring the northeast maple leaf powder into a new tube if the centrifugal tube has cracks; adding 700 mu L of mixed solution of 2 percent (g/ml) soluble pvp and 1 percent (mu L/ml) beta-mercaptoethanol with the same volume into a centrifugal tube containing plant tissue powder, shaking and uniformly mixing, putting ice on the ice for 10min, and centrifuging for 3min at 12500 rpm;
step three: discarding the supernatant, leaving the precipitate, adding 700 μ L of CTAB solution (containing RNaseA and 10 μ L of 10mg/ml RNaseA) preheated at 65 ℃ into a centrifugal tube containing the precipitate, shaking, uniformly mixing, putting into a 65 ℃ water bath kettle, carrying out water bath cracking for 10min, reversing and uniformly mixing every 5min, and centrifuging at 12500rpm for 5 min;
step four: leaving the supernatant, discarding the precipitate, transferring the supernatant into a new sterile 1.5ml centrifuge tube, adding 600. mu.L chloroform into the tube, shaking for 30s, and centrifuging at 12500rpm for 5 min;
step five: keeping the upper water phase, discarding the lower water phase, taking 400 μ L of the upper water phase to a new sterile 1.5ml centrifuge tube, adding 800 μ L of isoamyl alcohol into the tube, turning up and down gently, and centrifuging at 13500rpm for 15min after flocculent precipitate appears;
step six: discarding the supernatant, leaving a precipitate, adding 700 microliter of 70% ethanol into the centrifuge tube containing the precipitate, turning over and washing the precipitate for 30s, and repeating the step twice; pouring off ethanol in the tube, keeping the precipitate, centrifuging at 12500rpm for 2min, carefully sucking out the excessive liquid by using a gun head, and then airing the precipitate in a fume hood; dissolving the dried precipitate with 50 μ L sterilized deionized water to obtain the genomic DNA solution of Acer mono Maxim.
The extracted northeast maple DNA can be used for measuring concentration, detecting strips by agarose gel electrophoresis, PCR amplification and the like; the product can be stored in a refrigerator at 4 deg.C for short time, and stored at-30 deg.C or-80 deg.C for long time.
As shown in fig. 2, when the genomic DNA of acer northeast was extracted by the conventional CTAB method, the extracted DNA sample had many impurities, and even the extracted DNA was viscous and could not run out of the gel pore by electrophoresis.
As shown in FIG. 1, after the method of the invention is used for extracting genomic DNA of acer mono, the extracted DNA impurities are greatly reduced, the acer mono is not sticky any more, and the strips are clear; the extraction process is relatively simple, the steps are fewer, and the time is short; the cost of DNA extraction is also greatly reduced.
The above description is a preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical scope of the present invention, the technical solutions and the inventive concepts of the present invention with equivalent or modified alternatives and modifications within the technical scope of the present invention.
Claims (7)
1. A rapid extraction method of genomic DNA of acer northeast is characterized in that: the method comprises the following steps:
the method comprises the following steps: putting a sterilized steel ball into a sterilized 2ml centrifuge tube, cutting the acer palmatum leaves into pieces, adding the cut acer palmatum leaves into the centrifuge tube, putting the sterilized steel ball into the centrifuge tube, quickly putting the acer palmatum leaves into liquid nitrogen for freezing, and grinding the acer palmatum leaves into powder by a high-throughput tissue grinder;
step two: checking whether the centrifugal tube has cracks or not, and transferring the northeast maple leaf powder into a new tube if the centrifugal tube has cracks; adding a mixed solution of 2% (g/ml) soluble PVP and 1% (mu L/ml) beta-mercaptoethanol with the same volume into the powder, shaking and uniformly mixing, carrying out ice bath for 10min, centrifuging, and then removing supernate to obtain a precipitate;
step three: discarding the supernatant, leaving a precipitate, adding a prepared CTAB extraction buffer solution containing RNaseA into the precipitate, shaking and uniformly mixing, putting into a 65 ℃ water bath kettle, carrying out water bath cracking for 10min, and centrifuging to obtain a supernatant;
step four: the supernatant is left, the precipitate is discarded, chloroform is added into the supernatant, and the supernatant is centrifuged after shaking and mixing to obtain an upper aqueous phase;
step five: keeping the upper aqueous phase, discarding the lower aqueous phase, adding isoamylol into the upper aqueous phase, uniformly mixing and centrifuging to obtain a precipitate;
step six: discarding the supernatant, leaving a precipitate, washing the precipitate with 70% ethanol twice, air-drying the ethanol, and dissolving the precipitate with deionized water to obtain the genomic DNA solution of the acer mono.
2. The method for rapidly extracting genomic DNA of acer northeast as claimed in claim 1, wherein: the northeast maple leaves selected in the step one are healthy green leaves, the petioles are removed when the leaves are cut into pieces, the weight is 1g-2g, and the high-throughput tissue grinder is set to be ground for 2min at 65 Hz.
3. The method for rapidly extracting genomic DNA of acer northeast as claimed in claim 1, wherein: and in the second step, 700 mu L of mixed solution of 2 percent (g/ml) of soluble PVP and 1 percent (mu L/ml) of beta-mercaptoethanol with the same volume is added, ice bath is performed on the mixed solution, and the mixed solution is centrifuged at 12500rpm for 3 min.
4. The method for rapidly extracting genomic DNA of acer northeast as claimed in claim 1, wherein: in the third step, CTAB extraction buffer solution is preheated CTAB solution 700 mu L, and 1ml of CTAB is added into 10 mu L of RNaseA of 10 mg/ml; turning upside down and mixing once every 5min while water bath; centrifuging at 12500rpm for 5min at a temperature of 15 deg.C.
5. The method for rapidly extracting genomic DNA of acer northeast as claimed in claim 1, wherein: and step four, specifically, transferring the supernatant into a new sterile 1.5ml centrifuge tube, adding 600 mu L of chloroform into the centrifuge tube, shaking for 30s, and centrifuging for 5min at 12500 rpm.
6. The method for rapidly extracting genomic DNA of acer northeast as claimed in claim 1, wherein: and step five, specifically, taking 400 mu L of the upper-layer water phase to a new sterile 1.5ml centrifugal tube, adding 800 mu L of isoamyl alcohol into the centrifugal tube, slightly turning the centrifugal tube upside down, and centrifuging the centrifugal tube at 13500rpm for 15min after flocculent precipitates appear.
7. The method for rapidly extracting genomic DNA of acer northeast as claimed in claim 1, wherein: step six is specifically to add 700 μ L of 70% ethanol into the centrifuge tube containing the precipitate, turn over and wash the precipitate for 30s, and repeat the step twice; pouring out excessive ethanol in the tube, keeping precipitate, centrifuging at 12500rpm for 2min, sucking out excessive liquid with a gun head, and air drying in a fume hood; dissolving the dried precipitate with 50 μ L sterilized deionized water to obtain genomic DNA solution of Acer mono Maxim.
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| AU2020102458A4 (en) * | 2020-09-28 | 2020-11-12 | Research Institute of Subtropical Forestry, Chinese Academy of Forestry | Method for extracting high-quality dna from idesia polycarpa |
| CN113502286A (en) * | 2021-08-16 | 2021-10-15 | 天津诺禾致源生物信息科技有限公司 | Method for extracting DNA from plant tissue rich in secondary metabolites |
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- 2022-03-14 CN CN202210246732.3A patent/CN114507661A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101914522A (en) * | 2010-01-30 | 2010-12-15 | 浙江省海洋水产养殖研究所 | Extraction method of mangrove plant DNA |
| CN105368815A (en) * | 2015-11-25 | 2016-03-02 | 上海派森诺生物科技股份有限公司 | Extracting method of polysaccharide and polyphenol plant genomes |
| CN105713902A (en) * | 2016-04-14 | 2016-06-29 | 中国科学院寒区旱区环境与工程研究所 | Method for extracting total DNA (deoxyribonucleic acid) from eremophytes |
| CN107699561A (en) * | 2017-12-07 | 2018-02-16 | 山西省农业科学院棉花研究所 | A kind of modified CTAB method extracts cotton genomic dna |
| AU2020102458A4 (en) * | 2020-09-28 | 2020-11-12 | Research Institute of Subtropical Forestry, Chinese Academy of Forestry | Method for extracting high-quality dna from idesia polycarpa |
| CN113502286A (en) * | 2021-08-16 | 2021-10-15 | 天津诺禾致源生物信息科技有限公司 | Method for extracting DNA from plant tissue rich in secondary metabolites |
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