CN102424822B - Method for extracting genomic DNA from young tung oil tree leaves - Google Patents
Method for extracting genomic DNA from young tung oil tree leaves Download PDFInfo
- Publication number
- CN102424822B CN102424822B CN 201110453531 CN201110453531A CN102424822B CN 102424822 B CN102424822 B CN 102424822B CN 201110453531 CN201110453531 CN 201110453531 CN 201110453531 A CN201110453531 A CN 201110453531A CN 102424822 B CN102424822 B CN 102424822B
- Authority
- CN
- China
- Prior art keywords
- dna
- genomic dna
- tung oil
- oil tree
- extraction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for extracting genomic DNA from young tung oil tree leaves, relating to the separation and extraction technology of genomic DNA from plants in the molecular biology experiments. The method mainly comprises the steps of leaf tissue sample pretreatment, cell disruption, non-DNA component extraction, DNA precipitation, ion removal and tissue sample cyclic extraction. The method has the following advantages and positive effects: proteins, polyphenols, polysaccharides and other secondary metabolites in which the leaves are rich can be effectively removed by using washing liquid to pretreat the tissue samples of the young tung oil tree leaves three times; the step of tissue sample cyclic extraction is absent in the traditional methods for extracting genomic DNA; one tissue sample can undergo genomic DNA extraction four times, thus obviously increasing the yield of the genomic DNA; and the genomic DNA with high yield and high quality can be extracted from the young leaves stored at low temperature for a long time and from fresh young leaves acquired at different growth temperatures by the method. The method is suitable for extraction of genomic DNA from young tung oil tree leaves.
Description
Technical field
The present invention relates to plant genome DNA separation and extraction technology in the molecular biology experiment, relate in particular to a kind of method for extracting genomic DNA from young tung oil tree leaves.
Background technology
The extraction of genomic dna is a very important link of molecular biology experiment.The Quality and yield of genomic dna directly affects the carrying out of subsequent experimental relevant with DNA in the molecular biosciences experiment.Medicinal plant, aromatic series plant and xylophyta are rich in secondary metabolite; To the plant that this class is rich in secondary metabolite, traditional DNA extraction method is difficult for mentioning high-quality genomic dna.The kind of secondary metabolite is extremely complicated in this class plant leaf, and Various Seasonal between different plant species even between same species, and the heterogeneous degree of secondary metabolite is all higher between the blade of different leaf ages; Therefore, the genome DNA extracting method of a kind of plant often can not be advantageously applied to the extraction of another kind of plant genome DNA.
Tung oil tree originates in China, has high industrial value.The tung oil that extracts from tung oil tree paulownia fruit is the traditional large foreign exchange earning goods and materials of China.In China, tung oil is mainly used in the production of printing ink and coating.Now, energy dilemma is constantly aggravated, and environmental problem is day by day serious, and is extremely urgent to the demand of continuable new cleaning fuel.Tung oil tree is chosen as the energy-source plant species, and the tung oil that extracts from the tung oil tree fruit can be used as the raw material of production biofuel.Therefore will and tung oil be converted into biofuel to the breeding high-yield kind with the advanced deep research tung oil tree resource of molecular biology method theoretical foundation and technical support will be provided.The at present research of tung oil tree resource molecule aspect also is in the starting stage, and the tung oil tree genomic dna that extracts the high quality high yield is the basis of carrying out a series of follow-up molecular biology experiments.The tung oil tree blade is rich in polysaccharide, tannin, albumen and polyphenols, and these secondary metabolites affect the extraction of tung oil tree genomic dna.And between species the heterogeneity of secondary metabolite to cause the extracting method of other species gene group DNA be not the optimum extracting method of tung oil tree genomic dna.Research finds that the leaf age of tung oil tree blade affects the extraction of tung oil tree genomic dna.
Through retrieval, up to the present, also there is not special report for method for extracting genomic DNA from young tung oil tree leaves in the prior art.
Summary of the invention
Purpose of the present invention just is to overcome the shortcoming and defect that prior art exists, and is rich in the characteristics of polysaccharide, tannin, polyphenol and albumen for the tung oil tree spire, has proposed a kind of method for extracting genomic DNA from young tung oil tree leaves (abbreviation method).
The object of the present invention is achieved like this:
1, present method comprises the following steps:
1. with the spire tissue sample after wearing into fine powder under the liquid nitrogen flash freezer, get the bright leaf of 0.3g or 0.1g dry sample and change in the 2mL centrifuge tube;
2. add the 1.5mL washing lotion, behind the abundant mixing, static 15min in the ice, the centrifugal supernatant liquor of abandoning of 12000rpm;
3. 2. repeating step after twice, adds the 3%CTAB extracting solution of 500 μ l preheatings, abundant mixing, 65 ℃ of water-bath 40min;
4. the centrifugal 15min of 12000rpm normal temperature, shift supernatant liquor in new 1.5mL centrifuge tube, 24: 1 the chloroform-primary isoamyl alcohol that adds 100 μ l, fully behind the mixing, the centrifugal 15min of normal temperature 12000rpm, extracting 1～2 time, the supernatant liquor after the extracting change in the new centrifuge tube, and what add two volumes is pre-chilled to-20 ℃ 100% alcohol in advance; Be positioned over-20 ℃ of refrigerator-freezer 30min, allow the DNA precipitation fully separate out, the DNA precipitation of separating out is transferred in the 1.5mL centrifuge tube that contains 1mL 70% alcohol, rinsing 1～2 time, each 30min, the essence of falling the dry wine is after DNA precipitation drying, with the T of 100 μ l
0.1E fully dissolves, and is 37 ℃ of 10 μ g/mLRNA enzymes digestion 30min with 1 μ l concentration, and the DNA sample that this process obtains is called 1
StDNA;
5. step is shifted the tissue sample of centrifuge tube bottom after the supernatant liquor in 4., the 3%CTAB extracting solution that continues to add 500 μ l extracts, and this tissue sample circulates altogether and extracts 4 times, and the 3 batches of DNA samples of getting back according to this are called 2
NdDNA, 3
RdDNA and 4
ThDNA.
Described 100% alcohol refers to pure spirituous solution.
Described 70% alcohol refers to that 70mL straight alcohol usefulness sterile purified water constant volume is to the spirituous solution of 100mL.
In the multiple batches of DNA sample that the present invention extracts, the not low-temperature storage spire 1 that (30-37 ℃) gathers under the high growth temperature
StDNA, 2
NdDNA, 3
RdDNA and 4
ThDNA output is higher, and concentration is roughly between 300～800ng/ μ l.
In the multiple batches of DNA sample that the present invention extracts, the spire 1 that gathers under permanent low-temperature storage and the low growth temperature
StDNA yields poorly, and purity difference can discard the supernatant liquor of centrifugal generation after the water-bath, directly carry out the extraction of 3 times follow-up DNA; And 2
NdDNA, 3
RdDNA, 4
ThDNA output is high, and concentration is roughly between 100～700ng/ μ l.
The composition of washing lotion used in the present invention is: 50mM Tris-HCl, and 5mM EDTA, the 350mM sorbyl alcohol, 2%PVP, 0.5% beta-mercaptoethanol, pH value are 8.0.
Described 2% refers to 2g/100mL, and described 0.5% refers to 0.5mL/100mL.
3%CTAB extracting solution composition used in the present invention is: 3%CTAB, and 100mM Tris-HCl, 20mM EDTA, 1.4M NaCl, 2%PVP, 0.5% beta-mercaptoethanol, the pH value is 8.0.
Described 3% refers to 3g/100mL, and described 2% refers to 2g/100mL,
Described 0.5% refers to 0.5mL/100mL.
T used in the present invention
0.1The E composition is: 10mM Tris-HCl, and 0.1mM EDTA, pH 8.0.
Principle of work: the secondary metabolite that is rich in the tung oil tree spire is divided into from or suppresses the extraction etc. of DNA with DNA in the DNA extraction process such as polysaccharide, tannin, polyphenol etc.Before with 3%CTAB extracting solution lysing cell, with the washing lotion leaf tissue sample that grinds of pre-treatment liquid nitrogen repeatedly, front and back co-processing 3 times can be removed the secondary metabolite that disturbs DNA to separate in a large number first.Pretreated spire organizes sample to add the 3%CTAB extracting solution at 65 ℃ of lower lysing cell, the supernatant of centrifugal transfer passes through chloroform again: the extraction of primary isoamyl alcohol, the precipitation of alcohol, obtain cotton-shaped DNA precipitation, and utilize at last 70% washing with alcohol DNA precipitation, obtain the 1st DNA precipitation, namely 1
StDNA.And the sample of organizing that is positioned at behind the centrifugal transfer supernatant bottom the centrifuge tube can continue to add the 3%CTAB extracting solution, carries out the DNA extraction of a new round, organizes according to this sample to circulate altogether and extracts the 3 batches of dna samples of getting back 4 times.The separable tung oil tree genomic dna to high quality (few polysaccharide, few polyphenol, few pigment and protein etc.), high yield (organizing sample to obtain 4 batches of DNA precipitations for 1 part) of the present invention satisfies molecular biology experiment to the requirement of DNA concentration and quality.
2, the evaluation of tung oil tree genomic dna
The tung oil tree genome DNA sample that present method is extracted can be through 0.8% agarose gel electrophoresis and its concentration of UV spectrophotometer measuring and quality.
3, the purposes of present method
Present method can be extracted the high-quality genomic dna of high yield from the bright sample of tung oil tree spire and dry sample.The tung oil tree spire that (15 ℃-25 ℃) gather under the tung oil tree spire of (70 ℃) and the low growth temperature under the permanent low-temperature storage situation, the leaf internalization studies minute change of generation matter, cause the genomic dna output very low (sepharose can't detect) that the 1st extraction obtains in four circulation extractions of present method, purity is poor (260/280＜1.8) also, and the follow-up genomic dna output that obtains for three times is higher, purity is good, thus organize sample only extract 1 time traditional method difficult from permanent low-temperature storage the tung oil tree spire and low growth temperature under extract high yield and high-quality genomic dna the tung oil tree spire that gathers; But present method can overcome this permanent low-temperature storage and low growth temperature to the impact of spire DNA extraction, and rear 3 extractions can obtain the high-quality genomic dna of high yield preferably, satisfy the molecular biology subsequent experimental to the requirement of DNA concentration and quality.
The present invention has following advantages and positively effect:
1. the washing lotion in present method, first finishing oil paulownia spire is organized sample 3 times, can effectively remove the secondary meta-bolitess such as the polyphenol that is rich in the blade, polysaccharide, tannin;
2. the step that circulation is extracted in present method is unexistent in traditional genome DNA extracting method, organizes sample can carry out extracting genome DNA 4 times for 1 part, obtains the genomic dna sample 4 times, and genomic dna output is significantly improved;
3. extract the genomic dna of high quality high yield the tung oil tree spire that (15 ℃-25 ℃) gather under present method spire that can store from long-term low temperature (70 ℃) and the low growth temperature.
The present invention is applicable to the extraction of tung oil tree spire genomic dna.
Description of drawings
Fig. 1 is the electrophoresis picture (November, about 20 ℃ of temperature, 10 genotypic compound samples) with the bright sample genomic dna of tung oil tree spire compound sample of the low-temperature epitaxy of present method extraction;
Among the figure: 1,2,3,4 be respectively 1
St, 2
Nd, 3
Rd, 4
ThThe electrophoresis picture of DNA arranges 2 repetitions.
Fig. 2 is the situation that figure one described blade silica dehydrator is processed, and 3 repetitions are set.
Fig. 3 is the electrophoresis picture (July, about 37 ℃ of temperature) of the bright sample genomic dna of 6 genotype spires of tung oil tree that gathers under the high growth temperature of extracting with present method;
Among the figure: 1,2,3,4 be respectively 1
St, 2
Nd, 3
Rd, 4
ThThe electrophoresis picture of DNA arranges 2 repetitions, and A, B, C, D, E, F represent 6 genotype.
Fig. 4 is the electrophoresis picture (July, about 37 ℃ of temperature, 10 genotypic compound samples) of the spire compound sample genomic dna that gathers under the high growth temperature in 1 year of low-temperature storage of extracting with present method;
Among the figure: 1,2,3,4 be respectively 1
St, 2
Nd, 3
Rd, 4
ThThe electrophoresis picture of DNA arranges 2 repetitions.
English to Chinese:
1, CTAB:Cetyltrimethyl Ammonium Bromide, cetyl trimethylammonium bromide;
2, EDTA:Ethylene Diamine Tetraacetic Acid, ethylenediamine tetraacetic acid (EDTA);
3, PVP:Polyvinyl Pyrrolidone, polyvinylpyrrolidone;
4, β-Mercaptoethanol: mercaptoethanol.
Embodiment
Describe in detail below in conjunction with drawings and Examples:
One, extracting method
Choose 3 parts of 2 parts in the bright sample of tung oil tree spire compound sample that growth temperature about 20 ℃ (November) gathers and dry samples, and 2 parts of the tung oil tree spire compound samples in 1 year of the low-temperature storage that gathers under 37 ℃ of growth temperatures of temperature and without each two parts in the bright sample of 6 genotype spires of the tung oil tree of low-temperature storage, use the inventive method to carry out following 19 times and extract operations:
1. in liquid nitrogen with the abundant grind into powder of leaf tissue sample, take by weighing rapidly the bright leaf of 0.3g, or the 0.1g dry sample, fast transfer is in the 2mL centrifuge tube.Put into-70 ℃ of refrigerators, stand-by;
2. add the 1.5mL washing lotion, behind the abundant mixing, static 15min in the ice, the centrifugal supernatant liquor of abandoning of 12000rpm;
3. 2. repeating step after twice, adds the 3%CTAB extracting solution of 500 μ l preheatings, abundant mixing, 65 ℃ of water-bath 40min;
4. the centrifugal 15min of 12000rpm normal temperature, shift supernatant liquor in new 1.5mL centrifuge tube, 24: 1 the chloroform-primary isoamyl alcohol that adds 100 μ l, fully behind the mixing, the centrifugal 15min of normal temperature 12000rpm, extracting 2 times, the supernatant liquor after the extracting change in the new centrifuge tube, and what add two volumes is pre-chilled to-20 ℃ 100% alcohol in advance; Be positioned over-20 ℃ of refrigerator-freezer 30min, allow the DNA precipitation fully separate out, the DNA precipitation of separating out is transferred in the 1.5mL centrifuge tube that contains 1mL 70% alcohol, rinsing 2 times, each 30min, the essence of falling the dry wine is after DNA precipitation drying, with the T of 100 μ l
0.1E fully dissolves, and is 37 ℃ of 10 μ g/mL RNA enzymes digestion 30min with 1 μ l concentration, and the DNA sample that this process obtains is called 1
StDNA;
5. step is shifted the tissue sample of centrifuge tube bottom after the supernatant liquor in 4., continue to add the CTAB extracting solution of preheating, abundant mixing carries out the DNA extraction step of a new round, and the DNA sample that obtains in this process is 2
NdDNA, circulation gets back 3 according to this
RdDNA and 4
ThDNA.
Two, agarose gel electrophoresis method detects the tung oil tree spire genomic dna that present method is extracted
Such as Fig. 1,4 DNA that present method is extracted the bright sample of about 20 ℃ tung oil tree spires that gather of 11 month temperatures, 1
StDNA does not detect band, 2 on agarose
Nd, 3
Rd, 4
ThDNA all detects clear and bright electrophoretic band.The extraction of the bright leaf genomic dna of tung oil tree spire of growing under the suitable low temperature of present method is described.
Such as Fig. 2,4 DNA that present method is extracted behind silica dehydrator about 20 ℃ tung oil tree spires that gather of 11 month temperatures, 1
StDNA does not detect band, 2 on agarose
Nd, 3
Rd, 4
ThDNA all detects clear and bright electrophoretic band.The extraction of the tung oil tree spire dry sample genomic dna of growing under the suitable low temperature of present method is described.
Such as Fig. 3,4 DNA that present method is extracted the bright sample of 6 genotype spires of tung oil tree without low-temperature storage from July, 1
St, 2
Nd, 3
Rd, 4
ThDNA all detects clear and bright electrophoretic band.Compare with Fig. 1, Fig. 2, difference is 6 genotypic 1
StDNA all detects clear bright electrophoretic band.Illustrate that secondary metabolite in the tung oil tree spire shows obvious heterogeneity with the reduction of growth temperature, the extraction of genomic dna is caused obvious impact.Further specify the extraction of the tung oil tree spire DNA that grows under the suitable differing temps of present method.
Such as Fig. 4, present method is to storing 4 batches of genomic dnas of the spire compound sample extraction in July in 1 year, 1
StDNA does not detect band on agarose, and 2
NdDNA, 3
RdDNA, 4
ThDNA all detects clear and bright electrophoretic band.Compare with Fig. 3, after difference is to store 1 year from the tung oil tree spire in July, 1
StDNA does not detect band on sepharose.Illustrate that the long-time storage of low temperature may cause that the secondary metabolite composition changes in the tung oil tree spire, with store before do not show obvious heterogeneity, the extraction of DNA is caused obvious impact.Further specify the extraction that present method is fit to the tung oil tree spire genomic dna under the long-time condition of storage of low temperature.
Fig. 1 to Fig. 4, analysis-by-synthesis can be seen if adopt traditional genome DNA extracting method, organizes sample only to carry out 1 time and extracts, the low tung oil tree spire that gathers of tung oil tree spire, growth temperature to permanent storage extracts genomic dna, and the genomic dna output that obtains may be very low.And use the inventive method, and can greatly improve the output of tung oil tree spire genomic dna, also can overcome under the varying environment between tung oil tree spire blade the chemical ingredients heterogeneity spire scope that is used for the tung oil tree extracting genome DNA has been widened in the impact of DNA extraction.
Three, ultraviolet spectrophotometer method detects the Quality and yield of the tung oil tree spire genomic dna of present method extraction
The 4 batches of genomic dnas of spire that gather under the low growth temperature of extracting take present method detect its Quality and yield as example with ultraviolet spectrophotometer (model is Ultrospec 1100).Measure respectively 4 batches of DNA at the light absorption value of wavelength 260 nanometers and 280 nanometers, and calculate A260/A280 ratio, the results are shown in following table:
1 of the tung oil tree genomic dna sample that obtains by present method
StDNA A260/A280 ratio is 1.61, illustrates 1
StDNA contains more impurity, and 2
NdDNA, 3
RdDNA and 4
ThDNA all within 1.8～1.9 scopes, illustrates that the impurity such as pigment, protein are less in the genomic dna sample that obtains, and genomic dna purity is higher.Except 1
StOutside the DNA, the concentration range of rear 3 batches of genomic dnas is between 300～750ng/ μ l, and the tung oil tree genomic dna output that obtains is higher.
Claims (1)
1. a method for extracting genomic DNA from young tung oil tree leaves is characterized in that comprising the following steps:
1. with the spire tissue sample of bright leaf or dry sample after wearing into fine powder under the liquid nitrogen flash freezer, get the bright leaf fine powder of 0.3g or 0.1g dry sample fine powder and change in the 2mL centrifuge tube;
2. add the 1.5mL washing lotion, behind the abundant mixing, static 15min in the ice, the centrifugal supernatant liquor of abandoning of 12000rpm;
3. 2. repeating step after twice, adds the 3%CTAB extracting solution of 500 μ l preheatings, abundant mixing, 65 ℃ of water-bath 40min;
4. the centrifugal 15min of 12000rpm normal temperature, shift supernatant liquor in new 1.5mL centrifuge tube, chloroform-the primary isoamyl alcohol that adds the 24:1 of 100 μ l, fully behind the mixing, the centrifugal 15min of normal temperature 12000rpm, extracting 1～2 time, the supernatant liquor after the extracting change in the new centrifuge tube, and what add two volumes is pre-chilled to-20 ℃ 100% alcohol in advance; Be positioned over-20 ℃ of refrigerator-freezer 30min, allow the DNA precipitation fully separate out, the DNA precipitation of separating out is transferred in the 1.5mL centrifuge tube that contains 1mL 70% alcohol, rinsing 1～2 time, each 30min, the essence of falling the dry wine is after DNA precipitation drying, with the T of 100 μ l
0.1E fully dissolves, and is 37 ℃ of 10 μ g/mLRNA enzymes digestion 30min with 1 μ l concentration, and the DNA sample that this process obtains is called 1
StDNA;
5. step is shifted the tissue sample of centrifuge tube bottom after the supernatant liquor in 4., the 3%CTAB extracting solution that continues to add 500 μ l extracts, and this tissue sample circulates altogether and extracts 4 times, and the 3 batches of DNA samples of getting back according to this are called 2
NdDNA, 3
RdDNA and 4
ThDNA;
The composition of described washing lotion is: 50mM Tris-HCl, and 5mM EDTA, the 350mM sorbyl alcohol, 2%PVP, 0.5% beta-mercaptoethanol, the pH value is 8.0;
Described 3%CTAB extracting solution composition is: 3%CTAB, and 100mM Tris-HCl, 20mM EDTA, 1.4MNaCl, 2%PVP, 0.5% beta-mercaptoethanol, the pH value is 8.0;
Described T
0.1The E composition is: 10mM Tris-HCl, and 0.1mM EDTA, pH 8.0.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110453531 CN102424822B (en) | 2011-12-30 | 2011-12-30 | Method for extracting genomic DNA from young tung oil tree leaves |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110453531 CN102424822B (en) | 2011-12-30 | 2011-12-30 | Method for extracting genomic DNA from young tung oil tree leaves |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102424822A CN102424822A (en) | 2012-04-25 |
| CN102424822B true CN102424822B (en) | 2013-01-09 |
Family
ID=45958852
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110453531 Expired - Fee Related CN102424822B (en) | 2011-12-30 | 2011-12-30 | Method for extracting genomic DNA from young tung oil tree leaves |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102424822B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103540584B (en) * | 2012-07-10 | 2016-05-18 | 山东省果树研究所 | Be used for the preparation method of sample of the genomic DNA that extracts test tube sapling |
| CN103421765A (en) * | 2013-08-02 | 2013-12-04 | 山东农业大学 | DNA extracting method for analyzing MSAP (Methylation Sensitive Amplification Polymorphism) of epiphylla |
| CN108410863B (en) * | 2018-05-28 | 2020-10-16 | 中南大学湘雅二医院 | Efficient extraction method of guava leaf genome DNA |
-
2011
- 2011-12-30 CN CN 201110453531 patent/CN102424822B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN102424822A (en) | 2012-04-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102424823B (en) | Method for extracting genomic DNA from mature and old tung oil tree leaves | |
| CN104178480B (en) | Using the kit and method of DNA adsorption column rapid extraction DNA of plants | |
| CN102424822B (en) | Method for extracting genomic DNA from young tung oil tree leaves | |
| CN102206626A (en) | Method for extracting total RNA of multifarious fruit trees | |
| CN102443564B (en) | Method for extracting nucleuses of pear pollen tube | |
| CN101182344A (en) | Method for extracting and purifying DNA of plants like cactus | |
| CN108849498B (en) | Method for promoting accumulation of flavonoid substances in leaf tissues of ornamental crabapple | |
| CN102533728A (en) | Method for extracting high-quality cell nucleus DNA (Deoxyribonucleic Acid) of plant rich in polysaccharide and polyphenol | |
| CN101899435A (en) | High-throughput extraction method for sugarcane leaf genome by using ball mill | |
| CN105925567A (en) | Efficient and stable fruit tree RNA extraction method | |
| CN105087636A (en) | Tomato fruit gene transformation method based on agrobacterium injection | |
| CN103243088A (en) | Method for extracting high-quality DNA (deoxyribonucleic acid) in dry leaves of soybeans suitable for transgenic detection | |
| CN102899318B (en) | Method for extracting nanmu RNA (Ribonucleic Acid) | |
| CN102433324B (en) | Circular extraction method of plant genome DNA (deoxyribonucleic acid) | |
| CN102424825B (en) | Method for extracting total RNA from leaves and flower buds of filbert | |
| Linskens | Activation of the ovary | |
| CN103409410B (en) | Method for extracting the total microbial genome in vinegar fermentation process | |
| CN103266107B (en) | A kind of extracting method of arid biogeographic zone fruit tree crop genomic dna | |
| Lee et al. | Mechanism of release of nucleotidic material by fermenting brewer's yeast | |
| CN103540585A (en) | Method for extracting potato tuber total RNA (ribonucleic acid) by means of improved Trizol | |
| Hisamoto et al. | Protoplast isolation from bamboo leaves | |
| CN110577950A (en) | Extracting solution for extracting trace medicinal plant sample DNA and extraction method thereof | |
| CN104152561B (en) | A kind of authentication method of Kiwifruit nature heterozygote | |
| CN113444718B (en) | Method for extracting DNA from citrus medicinal material | |
| CN102816757A (en) | Method for obtaining high-quality fermented grain microorganism genome DNA (deoxyribonucleic acid) by pretreatment |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130109 Termination date: 20131230 |