CN102250882A - Method for extracting DNA (Deoxyribonucleic Acid) of total genome from zooplankter and intestinal inclusions thereof - Google Patents
Method for extracting DNA (Deoxyribonucleic Acid) of total genome from zooplankter and intestinal inclusions thereof Download PDFInfo
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- CN102250882A CN102250882A CN 201110175350 CN201110175350A CN102250882A CN 102250882 A CN102250882 A CN 102250882A CN 201110175350 CN201110175350 CN 201110175350 CN 201110175350 A CN201110175350 A CN 201110175350A CN 102250882 A CN102250882 A CN 102250882A
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Abstract
The invention discloses a method for extracting DNA (Deoxyribonucleic Acid) of a total genome from zooplankter and intestinal inclusions thereof. The method comprises the following steps of: immobilizing the zooplankter by utilizing a neutral Ruge's solution so that the intestinal inclusions are not lost and then to washing the body surface by utilizing sterile seawater filtered by a filtering membrane of 0.2 micron to remove other living beings attached on the surface so that polluting the zooplankter and the DNA of the intestinal inclusions is avoided; then washing by utilizing sterile super-pure water to reduce influence on DNA extraction caused by the salt ions in seawater; crushing and then cracking by utilizing a DNA cracking solution to release and dissociate the DNA of the zooplankter and intestinal inclusions to the cracking solution; then extracting the DNA of the total genome from the zooplankter and the intestinal inclusions thereof through utilizing a regular method. By using the method provided by the invention, the extraction of the DNA of the total genome from the zooplankter and the intestinal inclusions thereof is realized, and the loss of the DNA amount of the intestinal inclusions of the zooplankter is reduced to the maximum extent; and high-quality DNA samples for accurately constructing a qualitative structure of a spot plankton food web are provided, and the function and the efficiency of a regionally ecological system are further analyzed.
Description
Technical field:
The invention belongs to biological technical field, be specifically related to the extracting method of a kind of zooplankton and enteron aisle inclusion total genomic dna thereof.
Background technology:
Zooplankton is the secondary producer of food chain, is in the hub site of forming a connecting link in the food web of ocean, is the main driver of marine organisms pump, and is remarkable to the biomass geochemistry Circulation of source of students key element.Therefore, carry out its ecological study of ingesting to understanding the structure function of Marine ecosystems, quantize the cascade effect of matter and energy along food chain, it is significant to inquire into marine site biological production force level and Changing Pattern thereof.The utilization Protocols in Molecular Biology, can break through " cultivation " link, directly study the enteron aisle food of zooplankton in the physical environment and form and the efficient of ingesting, clear and definite its corresponding function reaction, accurately make up the qualitative framework of the on-the-spot food web that swims, and then the function of the resolution areas ecosystem and efficient.These researchs will be prerequisite to obtain high quality zooplankton enteron aisle inclusion genomic dna all.But because the zooplankton body surface often is attached with other biological, therefore to accurately obtain the zooplankton situation of ingesting, will guarantee that at first its body surface does not have other biological and pollutes; Obtain high-quality enteron aisle DNA sample, must guarantee that zooplankton historrhexis is complete, and be permitted to guarantee dna cleavage liquid tissue and the enteron aisle inclusion thereof of cracking zooplankton fully, thereby obtain high-quality DNA.
CTAB is a kind of cationic detergent, has the characteristic of precipitate nucleic acids; SDS makes lysis, and the histone that closely links to each other with the dna double chain is separated and sex change; EDTA chelating Mg
2+Or Mn
2+Ion can suppress the activity of nuclease; The Proteinase K protein hydrolysate, stable in SDS, be not subjected to the inhibition of EDTA, temperature of reaction is generally 50-55 ℃, can be used to digest various albumen; Tri-HCl provides a buffer environment, prevents that nucleic acid is destroyed; NaCl provides a hypersaline environment, and nucleic acid is fully dissolved, and is present in the liquid phase.Impurity such as the protein of chloroform removal sex change and polysaccharide, lipid.
At present, the patent report that does not still have zooplankton and enteron aisle inclusion total genomic dna extracting method.
Summary of the invention:
The purpose of this invention is to provide and a kind ofly can from zooplankton, extract zooplankton and enteron aisle inclusion total genomic dna thereof, and can reduce the zooplankton of loss of enteron aisle inclusion DNA amount of zooplankton and the extracting method of enteron aisle inclusion total genomic dna thereof to greatest extent.
The extracting method of zooplankton of the present invention and enteron aisle inclusion total genomic dna thereof may further comprise the steps:
(a) zooplankton is liquid-solid fixed with neutral Lu Geshi, again the zooplankton that fixes of same kind is divided together, the zooplankton of the same kind that will fix does not have other biological until body surface and adheres to aseptic and wash the body surface of zooplankton repeatedly through the seawater of 0.2 μ m membrane filtration then; Take out zooplankton, clean with aseptic ultrapure water, take out water is blotted, freezing one-tenth piece grinds broken again;
(b) will grind broken zooplankton and use the cracking of dna cleavage liquid, 55 ℃ of cracking 2～48h, with the DNA in the solution after the ordinary method extraction cracking, resulting DNA is zooplankton and enteron aisle inclusion total genomic dna thereof then;
Described dna cleavage liquid is Tris-HCl 10～50mM, SDS 5～10g/L, and EDTA 100mM, Proteinase K 200～500 μ g/mL, all the other are water, pH8.0.
Zooplankton that extraction obtains and enteron aisle inclusion total genomic dna thereof can be by concentration and the purity of UV spectrophotometer measuring DNA, and that guarantee follow-up use is high-quality DNA.
In the described step (a) with the freezing one-tenth piece of zooplankton, preferably with liquid nitrogen or low temperature with its freezing one-tenth piece, more preferably freezing with very low temperature with-80 ℃, because the 1.5ml centrifuge tube words of liquid nitrogen freezing, inconvenience operates, and the danger in order to reduce a staff, preferentially select for use-80 freezing.
Described step (b) will be ground broken zooplankton and be used the cracking of dna cleavage liquid, and preferably at 55 ℃ of cracking 48h, described dna cleavage liquid is Tris-HCl 10mM, SDS 5g/L, and EDTA 100mM, Proteinase K 200 μ g/mL, all the other are water, pH8.0.This lysate not only can be good at cracking zooplankton tissue and enteron aisle inclusion thereof under this cracking condition, and its consumption is few, and does not need NaCl, and is therefore both economical.
Neutral Shandong of the present invention Grignard solution belongs to article of the prior art, and it prepares in accordance with the following methods: the prescription of the neutral Shandong Grignard solution of every 100ml: iodine 5g, potassiumiodide 10g and ultrapure water 100ml.Take by weighing iodine 5g and potassiumiodide 10g respectively, add in the 80ml ultrapure water, after the stirring and dissolving, be settled to 100ml.
The present invention is liquid-solid fixed with neutral Lu Geshi with zooplankton; the enteron aisle inclusion that can make zooplankton can emptying and loss; thereby protection enteron aisle inclusion is subjected to the loss of minimum; for the total genomic dna that extracts zooplankton and enteron aisle inclusion thereof is laid a good foundation, thereby can react the situation of the enteron aisle inclusion of zooplankton really.Neutral Shandong Grignard solution can be good at fixedly zooplankton and enteron aisle inclusion thereof and can not extract subsequent DNA impacting, and the inventor extracts DNA after with the formaldehyde fixed zooplankton again, found that DNA extraction is failed.
The present invention is with aseptic and softly wash the zooplankton body surface repeatedly through the seawater of 0.2 μ m membrane filtration, general communications centre is more than 3 times, mainly be to rinse out for the other biological that the zooplankton body surface is adhered to, use microscopy again, guarantee that body surface does not have other biological and adheres to, this is in order to prevent that the body surface settled organism from polluting the enteron aisle inclusion.Aseptic seawater is with 0.2 μ m membrane filtration, and degerming is not only in its effect, removes also that all comprise bacterium, little algae, particulate matter, chip etc. greater than the impurity of 0.2 μ m in the seawater, avoids causing interference to extracting DNA.
The present invention with the flushing of aseptic ultrapure water through zooplankton aseptic and that clean up through its biology that the seawater of 0.2 μ m membrane filtration adheres to body surface, be to be rinsed totally, to reduce in the sterile filtration seawater salt ion to the influence of DNA extraction in order to ensure zooplankton sterile filtration seawater on one's body.
The present invention with liquid nitrogen or low temperature with the freezing one-tenth piece of zooplankton, grind broken, its main purpose is for zooplankton tissue and enteron aisle inclusion thereof is fully broken, makes cell fully cracking in dna cleavage liquid, reduces owing to grind insufficient loss that causes DNA output.
Use dna cleavage liquid of the present invention, through suitable temperature and suitable time, effectively zooplankton after the cracking fragmentation and enteron aisle inclusion thereof make nucleic acid dissociate out, and then just can extract DNA from lysate through conventional method.SDS can make the karyomit(e) segregation at 55 ℃～65 ℃ following lysing cell, and protein denaturation discharges nucleic acid; Proteinase K is stable in SDS, and temperature of reaction is generally 50-55 ℃, can be used to digest various albumen; EDTA suppresses the activity of DNA enzyme, and Tri-HCl provides a buffer environment, prevents that nucleic acid is destroyed.Cracking temperature is 55 ℃, and the cracking time is 2～48 hours, and overlong time may cause damage to DNA, and the time is too short then to make lysis incomplete.
The present invention is according to the physiological characteristic of zooplankton, utilize that neutral Lu Geshi is liquid-solid to decide zooplankton, make the enteron aisle inclusion of zooplankton can not lose, utilize aseptic again, and the seawater through 0.2 μ m membrane filtration softly washes the zooplankton body surface repeatedly, remove the other biological that the zooplankton body surface adheres to, thereby avoid the pollution of other biological that body surface adheres to zooplankton and enteron aisle inclusion DNA thereof, and then utilize aseptic ultrapure water to clean zooplankton, thereby reduce the influence that salt ion may cause DNA extraction in the sterile filtration seawater, through after the sufficient fragmentation, with dna cleavage liquid of the present invention cracking, make the DNA release in zooplankton and the enteron aisle inclusion thereof be free in the lysate, and then through conventional method, from lysate, extract DNA, be the total genomic dna of zooplankton and enteron aisle inclusion thereof.
Therefore the realization of success of the present invention to the extraction of the total genomic dna of zooplankton and enteron aisle inclusion thereof, this method can reduce the loss of zooplankton enteron aisle inclusion DNA amount to greatest extent, be accurately to make up the qualitative framework of the on-the-spot food web that swims, and then the function of the resolution areas ecosystem and efficient provide high-quality enteron aisle inclusion DNA sample.
Description of drawings:
Fig. 1 is the electrophorogram that embodiment 1 usefulness method of the present invention is extracted inferior strong very wise water flea and enteron aisle inclusion total genomic dna thereof, wherein 1 is Marker, and 2 and 3 all is very wise by force water flea in Asia and the total genomic DNA of enteron aisle inclusion thereof that extracts by method of the present invention;
Fig. 2 is that the inferior strong very wise water flea and the enteron aisle inclusion total genomic dna thereof that extract with embodiment 1 usefulness method of the present invention are the electrophorogram that template is carried out the product of PCR, wherein 1 is Marker, 2 negative contrasts, the 3rd, be the electrophorogram that template is carried out the product of pcr amplification with strong very wise water flea in Asia and enteron aisle inclusion total genomic dna thereof, the 4th, positive control is to be the electrophorogram of template pcr amplification product with the Phaeodactylum tricornutum genomic dna;
Fig. 3 uses fat soft arrow worms (Flaccisagitta enflata) and the enteron aisle inclusion total genomic dna electrophorogram thereof that method of the present invention is extracted among the embodiment 2, wherein 1 is Marker, and 2 is fat soft arrow worms and enteron aisle inclusion total genomic dna thereof;
Fig. 4 is to be the electrophorogram that template is carried out the product of PCR with soft arrow worms of obesity (Flaccisagitta enflata) and enteron aisle inclusion total genomic dna thereof, wherein 1 is Marker, 2 negative contrasts, the 3rd, be the electrophorogram that template is carried out the product of pcr amplification with Sagitta enflata and enteron aisle inclusion total genomic dna thereof, the 4th, positive control is to be the electrophorogram of template pcr amplification product with the Phaeodactylum tricornutum genomic dna.
Embodiment:
Following examples are to further specify of the present invention, rather than limitation of the present invention.
Embodiment 1:
In March, 2011, drag in karang sea area, Sanya horizontal direction with zooplankton I type net and to get zooplankton, with zooplankton with neutral Lu Geshi liquid-solid fixed after, under Stereo microscope, carry out the zooplankton sorting with disposable plastic tube, identical type is put into 1.5ml sterilization centrifuge tube respectively, and neutral Lu Geshi is liquid-solid fixed, and 4 ℃ of preservations are standby.Getting the strong very wise water flea Eucalanus subcrassus in 20 Asias that fix is put in the culture dish of diameter 3.5cm, with softly washing the zooplankton body surface repeatedly through the sterilization seawater behind the 0.2 μ m Millipore membrane filtration, microscopy guarantees that body surface does not have other biological and adheres to; Take out zooplankton, clean with the sterilization ultrapure water, salt ion is transferred to zooplankton in the 1.5ml centrifuge tube, and water is blotted the influence of DNA extraction in the sterile filtration seawater to reduce.Centrifuge tube is placed-80 ℃ of Ultralow Temperature Freezers freezing, take out behind the 10min,, repeat above operation 3 times immediately with the pestle broken 1-2min that mills; Adding 400 μ l dna cleavage liquid in centrifuge tube (10mMTris-HCl pH 8.0,5g/L SDS, 100mM EDTA pH8.0,200 μ gmL-1 proteinase K, all the other are water, pH8.0), the attached crop with pestle carries out wash-out simultaneously.Then at 55 ℃ of water-bath 48h, during jog centrifuge tube 3-5 time; In centrifuge tube, add 66 μ l 5MNaCl solution; The CTAB solution that adds 10% (the quality volume fraction) of 66 μ l after 56 ℃ of preheatings behind the mixing again, behind the mixing that vibrates on the whirlpool instrument, 56 ℃ of water-bath 10min; Add isopyknic chloroform then, mixing, the centrifugal 10min of 12000rpm; Supernatant is transferred in the new 1.5ml centrifuge tube; Carry out purifying (according to the operation of its specification sheets) with Zymo DNA concentrator (available from ZYMO RESEARCH company, article No. is No:D4034) test kit, with the DNA behind the purifying be dissolved in 50 μ lTris-HCl (10mM, pH8) in ,-20 ℃ of preservations.Get 4 μ lDNA and carry out agarose electrophoresis, its electrophorogram wherein is that present embodiment extracts the DNA that obtains in 2 and 3 the point sample hole as shown in Figure 1, this shows, can extract from zooplankton and enteron aisle inclusion thereof by method of the present invention and obtain DNA.
With the DNA that extracts is template, with 18S rDNA universal primer
(18SrDNA?comF1：GCTTGTCTCAAAGATTAAGCCATGC；
18SrDNA comR1:CACCTACGGAAACCTTGTTACGAC), sequence is respectively shown in SEQ ID NO.1 and SEQ ID NO.2.
The DNA that extraction is obtained increases, and amplification system is 25 μ l:17.375 μ l ddH
2O; 2.5 μ l Taq buffer; 2 μ ldNTP; 0.125 μ l ExTaq; Each 1 μ l of primers F 1 and R1; Template DNA 1 μ l.Amplification condition is: 94 ℃ of 3min; 5 * (94 ℃ of 20s; 52 ℃ of 30s; 72 ℃ of 1min); 30 * (94 ℃ of 20s; 56 ℃ of 30s; 72 ℃ of 1min); 72 ℃ of 7min; 15 ℃ of forever; Amplified fragments is 1.8kb.Get 4 μ l amplified productions and carry out electrophoresis, the result as shown in Figure 2, wherein 2 negative contrasts, the 3rd, be the electrophorogram that template is carried out pcr amplification product with strong very wise water flea in Asia and enteron aisle inclusion total genomic dna thereof, the 4th, positive control, be to be template with the Phaeodactylum tricornutum genomic dna, after increasing with above-mentioned primer and system, the electrophorogram of the product of amplification.As can be seen from the figure, can successfully extract with the method for present embodiment and obtain very wise by force water flea in Asia and enteron aisle inclusion total genomic dna thereof.
Embodiment 2:
In May, 2011, drag in Daya Gulf sea area horizontal direction with zooplankton I type net and to get zooplankton, with zooplankton with neutral Lu Geshi liquid-solid fixed after, under Stereo microscope, carry out the zooplankton sorting with disposable plastic tube, identical type is put into 1.5ml sterilization centrifuge tube respectively, and neutral Lu Geshi is liquid-solid fixed, and 4 ℃ of preservations are standby.Getting 20 the soft arrow worms of obesity (Flaccisagitta enflata) that fix is put in the culture dish of diameter 3.5cm, with softly washing the zooplankton body surface repeatedly through the sterilization seawater behind the 0.2 μ m Millipore membrane filtration, microscopy guarantees that body surface does not have other biological and adheres to; Take out zooplankton, clean with the sterilization ultrapure water, salt ion is transferred to zooplankton in the 1.5ml centrifuge tube, and water is blotted the influence of DNA extraction in the sterile filtration seawater to reduce.Centrifuge tube is placed-80 ℃ of Ultralow Temperature Freezers freezing, take out behind the 10min,, repeat above operation 3 times immediately with the pestle broken 1-2min that mills; Adding 500 μ l dna cleavage liquid in centrifuge tube (50mM Tris-HCl pH8.0,10g/L SDS, 100mM EDTApH8.0,500 μ gmL-1proteinase K, all the other are water, pH8.0), the attached crop with pestle carries out wash-out simultaneously.Then at 55 ℃ of water-bath 2h, during jog centrifuge tube 3-5 time; Behind the cooling 2min, add chloroform-primary isoamyl alcohol (24: 1) to full packages, jog 2～3min mixes both; The centrifugal 10min of 10000rpm meanwhile, adds the Virahol of 600 μ l in another new sterilization centrifuge tube; Supernatant is changed in the centrifuge tube that contains Virahol,, Virahol and water layer are fully mixed to see the DNA floss the centrifuge tube 30s that slowly fluctuates; Behind the centrifugal 1min of 10000rpm, outwell liquid immediately, note white DNA precipitation not being poured out, centrifuge tube is stood upside down on the paper handkerchief that spreads out; Behind the 60s, upright centrifuge tube adds the ethanol of volume fraction 75% of 720 μ l and the sodium-acetate of 80 μ l5M, rotates gently, with finger bomb tube point, the DNA block at precipitation and the pipe end is swum in liquid; Place 30min, make the impurity dissolving of DNA block; Behind the centrifugal 1min of 10000rpm, outwell liquid, add the ethanol of 800 μ l volume fractions 75% again, DNA is washed 30min again; Behind the centrifugal 30s of 10000rpm, outwell liquid immediately, centrifuge tube is stood upside down on the paper handkerchief that spreads out; After several minutes, upright centrifuge tube, dry DNA (natural air drying or dry up with air duct); Add 50 μ l, 0.5 * TE (containing RNase) damping fluid, make the DNA dissolving, place 37 ℃ of about 15h of thermostat container, RNA is cleared up;-20 ℃ of preservations.Get 4 μ lDNA and carry out agarose electrophoresis, its electrophorogram wherein is that present embodiment extracts the DNA that obtains in the 2 point sample holes as shown in Figure 3, this shows, can extract from zooplankton and enteron aisle inclusion thereof by method of the present invention and obtain DNA.
With 18S rDNA universal primer (18SrDNA comF1:GCTTGTCTCAAAGATTAAGCCATGC; 18SrDNA comR1:CACCTACGGAAACCTTGTTACGAC) DNA that extraction is obtained increases, and amplification system is 25 μ l:17.375 μ l ddH
2O; 2.5 μ l Taq buffer; 2 μ l dNTP; 0.125 μ l ExTaq; Each 1 μ l of primers F 1 and R1; Template DNA 1 μ l.Amplification condition is: 94 ℃ of 3min; 5 * (94 ℃ of 20s; 52 ℃ of 30s; 72 ℃ of 1min); 30 * (94 ℃ of 20s; 56 ℃ of 30s; 72 ℃ of 1min); 72 ℃ of 7min; 15 ℃ of forever; The amplified fragments theory is 1.8bp, get 4 μ l amplified productions and carry out electrophoresis, the result as shown in Figure 4, wherein 2 negative contrasts, the 3rd, be the electrophorogram that template is carried out pcr amplification product with Sagitta enflata and enteron aisle inclusion total genomic dna thereof, the 4th, positive control is to be template through the electrophorogram of the product of above-mentioned 18s rDNA universal primer amplification with the Phaeodactylum tricornutum genomic dna.As can be seen from the figure, can successfully extract with the method for present embodiment and obtain Sagitta enflata and enteron aisle inclusion total genomic dna thereof.
Claims (4)
1. the extracting method of zooplankton and enteron aisle inclusion total genomic dna thereof is characterized in that, may further comprise the steps:
(a) zooplankton is liquid-solid fixed with neutral Lu Geshi, again the zooplankton that fixes of same kind is divided together, the zooplankton of the same kind that will fix does not have other biological until body surface and adheres to aseptic and wash the body surface of zooplankton repeatedly through the seawater of 0.2 μ m membrane filtration then; Take out zooplankton, clean with aseptic ultrapure water, take out water is blotted, freezing one-tenth piece grinds broken again;
(b) will grind broken zooplankton and use the cracking of dna cleavage liquid, 55 ℃ of cracking 2～48h, with the DNA in the solution after the ordinary method extraction cracking, resulting DNA is zooplankton and enteron aisle inclusion total genomic dna thereof then;
Described dna cleavage liquid is Tris-HCl 10～50mM, SDS 5～10g/L, and EDTA 100mM, Proteinase K 200～500 μ g/mL, all the other are water, pH8.0.
2. the extracting method of zooplankton according to claim 1 and enteron aisle inclusion total genomic dna thereof is characterized in that, with the freezing one-tenth piece of zooplankton, is with its freezing one-tenth piece with liquid nitrogen or low temperature in the described step (a).
3. the extracting method of zooplankton according to claim 2 and enteron aisle inclusion total genomic dna thereof is characterized in that, described is freezing with-80 ℃ very low temperature with low temperature with the freezing one-tenth piece of zooplankton.
4. the extracting method of zooplankton according to claim 1 and enteron aisle inclusion total genomic dna thereof, it is characterized in that, described step (b) will be ground broken zooplankton and be used the cracking of dna cleavage liquid, be at 55 ℃ of cracking 48h, described dna cleavage liquid is Tris-HCl 10mM, SDS 5g/L, EDTA 100mM, Proteinase K 200 μ g/mL, all the other are water, pH8.0.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107144488A (en) * | 2017-03-14 | 2017-09-08 | 暨南大学 | A kind of fish gastrointestinal content quick quantitative analytic method and its application |
| CN109337955A (en) * | 2018-12-07 | 2019-02-15 | 暨南大学 | A kind of third generation sequencing zooplankter genome DNA extracting method and application |
| CN117126843A (en) * | 2023-09-18 | 2023-11-28 | 生态环境部华南环境科学研究所(生态环境部生态环境应急研究所) | DNA extraction method for small zooplankton single body |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000034504A2 (en) * | 1998-12-10 | 2000-06-15 | Genetic Vectors, Inc. | Method of protein removal |
| CN101696408A (en) * | 2009-10-23 | 2010-04-21 | 厦门大学 | Method for extracting DNA from single eggs of copepods |
| WO2011003184A1 (en) * | 2009-07-08 | 2011-01-13 | National Research Council Of Canada | Detection of microcystin-producing cyanobacteria |
-
2011
- 2011-06-27 CN CN 201110175350 patent/CN102250882B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000034504A2 (en) * | 1998-12-10 | 2000-06-15 | Genetic Vectors, Inc. | Method of protein removal |
| US20010049133A1 (en) * | 1998-12-10 | 2001-12-06 | Genetic Vectors, Inc. | Method of protein removal |
| WO2011003184A1 (en) * | 2009-07-08 | 2011-01-13 | National Research Council Of Canada | Detection of microcystin-producing cyanobacteria |
| CN101696408A (en) * | 2009-10-23 | 2010-04-21 | 厦门大学 | Method for extracting DNA from single eggs of copepods |
Non-Patent Citations (3)
| Title |
|---|
| 《Molecular Phylogenetics and Evolution》 20100301 Grace A. Wyngaard等 Phylogeny of the freshwater copepod Mesocyclops (Crustacea: Cyclopidae) based on combined molecular and morphological data, with notes on biogeography 2010 Elsevier Inc. 第753-764页 1-4 第55卷, * |
| 《厦门大学学报（自然科学版）》 20050531 周美玉等 甲醛溶液保存的中华哲水蚤基因组DNA提取和PCR扩增 第403-406页 1-4 第44卷, 第3期 * |
| 《海洋科学》 20051231 方旅平等 不同保存条件下中华哲水蚤基因组DNA提取的比较 第1-4页 1-4 第29卷, 第2期 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107144488A (en) * | 2017-03-14 | 2017-09-08 | 暨南大学 | A kind of fish gastrointestinal content quick quantitative analytic method and its application |
| CN107144488B (en) * | 2017-03-14 | 2019-06-21 | 暨南大学 | A kind of fish gastrointestinal content quick quantitative analytic method and its application |
| CN109337955A (en) * | 2018-12-07 | 2019-02-15 | 暨南大学 | A kind of third generation sequencing zooplankter genome DNA extracting method and application |
| CN109337955B (en) * | 2018-12-07 | 2021-12-03 | 暨南大学 | Third-generation sequencing zooplankton genome DNA extraction method and application |
| CN117126843A (en) * | 2023-09-18 | 2023-11-28 | 生态环境部华南环境科学研究所(生态环境部生态环境应急研究所) | DNA extraction method for small zooplankton single body |
| CN117126843B (en) * | 2023-09-18 | 2024-05-14 | 生态环境部华南环境科学研究所(生态环境部生态环境应急研究所) | DNA extraction method for small zooplankton single body |
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