CN109100335A - The method that a kind of pair of salmon trout embryo's living body carries out Ploidy Identification - Google Patents
The method that a kind of pair of salmon trout embryo's living body carries out Ploidy Identification Download PDFInfo
- Publication number
- CN109100335A CN109100335A CN201810631986.0A CN201810631986A CN109100335A CN 109100335 A CN109100335 A CN 109100335A CN 201810631986 A CN201810631986 A CN 201810631986A CN 109100335 A CN109100335 A CN 109100335A
- Authority
- CN
- China
- Prior art keywords
- embryo
- sample
- ploidy
- salmon trout
- living body
- 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.)
- Pending
Links
- 210000001161 mammalian embryo Anatomy 0.000 title claims abstract description 36
- 238000000034 method Methods 0.000 title claims abstract description 36
- 241000277269 Oncorhynchus masou Species 0.000 title claims abstract description 29
- 238000001514 detection method Methods 0.000 claims abstract description 17
- 210000002308 embryonic cell Anatomy 0.000 claims abstract description 12
- 238000012360 testing method Methods 0.000 claims abstract description 11
- 239000006285 cell suspension Substances 0.000 claims abstract description 10
- 238000004043 dyeing Methods 0.000 claims abstract description 10
- 238000004458 analytical method Methods 0.000 claims abstract description 4
- 102000002322 Egg Proteins Human genes 0.000 claims description 6
- 108010000912 Egg Proteins Proteins 0.000 claims description 6
- 210000004681 ovum Anatomy 0.000 claims description 6
- 239000007853 buffer solution Substances 0.000 claims description 3
- 210000004027 cell Anatomy 0.000 claims description 3
- 239000000706 filtrate Substances 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 239000000725 suspension Substances 0.000 claims description 3
- 238000003860 storage Methods 0.000 claims description 2
- 210000003754 fetus Anatomy 0.000 claims 1
- 238000007689 inspection Methods 0.000 claims 1
- 235000019688 fish Nutrition 0.000 abstract description 17
- 241000251468 Actinopterygii Species 0.000 abstract description 16
- 238000004519 manufacturing process Methods 0.000 abstract description 10
- 238000009395 breeding Methods 0.000 abstract description 9
- 230000001488 breeding effect Effects 0.000 abstract description 9
- 239000000975 dye Substances 0.000 description 13
- 210000004940 nucleus Anatomy 0.000 description 13
- 208000020584 Polyploidy Diseases 0.000 description 9
- 238000011160 research Methods 0.000 description 9
- 210000000349 chromosome Anatomy 0.000 description 6
- 241000277275 Oncorhynchus mykiss Species 0.000 description 5
- 208000026487 Triploidy Diseases 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 230000006698 induction Effects 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 3
- 230000000366 juvenile effect Effects 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 208000002109 Argyria Diseases 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 210000003743 erythrocyte Anatomy 0.000 description 2
- 238000000105 evaporative light scattering detection Methods 0.000 description 2
- 238000011031 large-scale manufacturing process Methods 0.000 description 2
- 210000002305 nucleolus organizer region Anatomy 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241001247278 Acanthopagrus schlegelii Species 0.000 description 1
- 241000972773 Aulopiformes Species 0.000 description 1
- 241001609213 Carassius carassius Species 0.000 description 1
- 241001275890 Megalobrama amblycephala Species 0.000 description 1
- 241000277326 Oncorhynchus gorbuscha Species 0.000 description 1
- 241001327684 Oncorhynchus mykiss aguabonita Species 0.000 description 1
- 241001280377 Oncorhynchus tshawytscha Species 0.000 description 1
- 241000276701 Oreochromis mossambicus Species 0.000 description 1
- 241000269811 Pagrus pagrus Species 0.000 description 1
- 241000694873 Paralichthyidae Species 0.000 description 1
- 241000269908 Platichthys flesus Species 0.000 description 1
- 241000277263 Salmo Species 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 230000005669 field effect Effects 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 230000031864 metaphase Effects 0.000 description 1
- 239000003226 mitogen Substances 0.000 description 1
- 230000011278 mitosis Effects 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Pathology (AREA)
- Physics & Mathematics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The method that a kind of pair of salmon trout embryo's living body carries out Ploidy Identification, it is related to the method that a kind of couple of fish embryo carries out Ploidy Identification.The method of the present invention can shorten the Ploidy Identification period, avoid production loss caused by extraneous factor in breeding process.Identification method: one, 5~8 DEG C under the conditions of salmon trout eyespot phase embryo is individually temporarily put, and keeps embryo's air exchange not anoxic；Two, embryonic cell suspension is made；Three, it filters, obtains detection sample；Four, the known salmon trout diploid sample of dyeing is detected with the ploidy analyser, and adjusts fluorescence signal value to being set as check sample at 100；It is detected after will test sample dyeing with the ploidy analyser, records fluorescence signal value, the analysis system carried by machine will test the peak value figure mean value of sample compared with check sample peak value figure, determine the ploidy size of detection sample.
Description
Technical field
The present invention relates to the methods that a kind of couple of fish embryo carries out Ploidy Identification.
Background technique
Fishes Chromosomes operating technology (gynogenesis and multiploid induction) plays important in aquatic livestock breeding field
Effect.Currently, the research work in relation to multiploid induction makes much progress in the world.The seas such as salmon trout class, flounder flounder class, Tilapia mossambica,
Triploid is induced in freshwater fish, in salmon fishes oneself obtain Atlantic salmon, rainbow trout (Salmo gairdneri), rainbow trout, hump back salmon,
The autotriploids such as king salmon and America red dot.It is domestic in recent years also successfully to have induced triploid megalobrama amblycephala, water
Brilliant coloured silk crucian carp, red porgy, black porgy, lefteye flounder and rainbow trout, especially in last century the eighties mid-term to the nineties initial stage, China Water obstetrics
It learns research institute Heilungkiang aquatic products research institute and has just carried out rainbow trout female Cultivating techniques research entirely, and the beginning of this century completes three times of rainbow trout
System kind technical research, tentatively realizes the large-scale production of triploid；Later, China Aquatic Science Research Institute Heilungkiang aquatic products
Research institute has also carried out the research of triploid Oncorhychus masou and triploid golden trout seeding technique.
There are many kinds of related fish methods for ploidy determination, comprising: chromosome counting method, red blood cell and nucleus volume measurement
Method, nucleolus organizer region's silver staining counting method, body cell DNA relative amount measuring method etc..Chromosome technique method needs to obtain Fish
Cell mitogen division phases (karyotype), by matching to chromosome, hand picking goes out all homologous dyes
Colour solid, and counted, and a kind of method for identifying the ploidy of fish is compared with reference genome, this method is current
Generally acknowledged most accurate fish methods for ploidy determination, but due to there are mitosis metaphase split coil method be prepared into power it is low with it is same
The artificial judgement of source chromosome pairing there are error probabilities high, level-one complex steps and it is professional too strong the features such as, be not suitable for carrying out
It is not very practical especially to carry out the screening of high-volume polyploid in production practice for mass detection.Red blood cell and nucleus volume
Mensuration and nucleolus organizer region's silver staining counting method measurement ploidy accuracy are lower, and after being not readily used for batch processing induction polyploid
The Ploidy Identification in generation.
When development Fishes Chromosomes double experiment, need effectively to assess set experiment condition, such as press
Power intensity, pressure effect time and after fertilization pressure intervention window phase etc.；And more times are being carried out for seed initiative enterprise
Also ploidy verifying is carried out to the polyploid seed of preparation in body scale production of hybrid seeds production, require a set of stabilization, quickly and just
Prompt methods for ploidy determination, especially in embryo stage.Because polyploid Seedling production enterprise will be while selling out seed
A times implementations are known at the first time.Currently, being all equal production of hybrid seeds offspring to the assessment of fish gynogenesis and multiploid induction result
It is developed to young stage and carries out Ploidy Identification again later, then assesses the superiority and inferiority that genome walking tests set parameter and condition.
Thus there are some defects: (1) since gynogenesis and multiploid induction technology offspring obtained must wait until to enter juvenile fish
Stage just can be carried out Ploidy Identification, and especially as salmon trout is long growth cycle, development to juvenile fish stage needs 6 months, this causes must
Palpus etc. is lot more time to the result of assessment genome walking experiment；(2) before due to assessment ploidy operation experiments result, have longer
One section of breeding process, in this process because caused by the reasons such as breeding environment variation, cultural technique and disease fish it is dead
Be it is unpredictable, the offspring of genome walking experiment preparation does not have equal detections, and just all dead cases also can be found everywhere.
Summary of the invention
In order to shorten the Ploidy Identification period, production loss caused by extraneous factor in breeding process is avoided, the present invention
Provide the method that a kind of pair of salmon trout embryo's living body carries out Ploidy Identification.
The method that the present invention carries out Ploidy Identification to salmon trout embryo's living body is as follows:
One, salmon trout eyespot phase embryo is individually temporarily put under the conditions of 5~8 DEG C, and keeps embryo's air exchange not anoxic；
Two, salmon trout eyespot phase embryo is pierced through with tip tweezers, releases the liquid fat in ovum, is pressed from both sides later with tweezers
Live in embryo stirs in PBS buffer solution and washes excess oil, is subsequently placed in the nucleus dye liquor of culture dish, by ovum skin and work
The chopping of body embryo tire shakes up embryonic cell suspension is made；
Three, it rinses culture dish with nucleus dye liquor to mix with step 2 embryonic cell suspension, then with 30 μm of aperture strainers
Embryonic cell mixing suspension is filtered；It takes filtrate to be refiltered once with 30 μm of aperture strainers, obtains detection sample；
Four, the known salmon trout diploid sample of dyeing is detected with the ploidy analyser, and adjusts fluorescence signal value to 100
Place is set as check sample；It is detected after will test sample dyeing with the ploidy analyser, records fluorescence signal value, it is included by machine
Analysis system, will test the peak value figure mean value of sample compared with check sample peak value figure, determine detection sample ploidy size.
The present invention carries out Ploidy detection and identification in embryo stage, can carry out without until offspring enters juvenile fish stage
Detection cycle is greatly shortened in Ploidy Identification, knows ploidy in fertilized eggs period, can do the production of hybrid seeds of polyploid again in due course.For
Aquaculture enterprise and research institution can carry out a large amount of and multiple batches of identifications in breeding period, due to the fish oosperm incubation period
It is longer, therefore obtain qualification result in embryo stage, breeding period is carried out again the preparation of multiple polyploid be it is extremely beneficial and
It is feasible.
The present invention is short to the pre-treatment, dyeing and detection time of test sample, easy to operate, quick, identification stability it is good,
Accuracy rate is high, dyeing drug dosage is small, the micro- poison of operation toxicity, to body fanout free region, be suitble to the mass detection of industrialization production
With identification, it is more conducive to polyploid breeding practice and polyploid large-scale production needs.The method of the present invention is universal for technology
Stronger with promoting, without profession, upper machine personnel are assisted, independent training i.e. detection operation in 20 minutes.
Specific embodiment
The technical solution of the present invention is not limited to the following list, further includes between each specific embodiment
Any combination.
Specific embodiment 1: the method that present embodiment carries out Ploidy Identification to salmon trout embryo's living body is as follows:
One, salmon trout eyespot phase embryo is individually temporarily put under the conditions of 5~8 DEG C, and keeps embryo's air exchange not anoxic；
Two, salmon trout eyespot phase embryo is pierced through with tip tweezers, releases the liquid fat in ovum, is pressed from both sides later with tweezers
Live in embryo stirs in PBS buffer solution and washes excess oil, is subsequently placed in the nucleus dye liquor of culture dish, by ovum skin and work
The chopping of body embryo tire shakes up embryonic cell suspension is made；
Three, it rinses culture dish with nucleus dye liquor to mix with step 2 embryonic cell suspension, then with 30 μm of aperture strainers
Embryonic cell mixing suspension is filtered；It takes filtrate to be refiltered once with 30 μm of aperture strainers, obtains detection sample；
Four, the known salmon trout diploid sample of dyeing is detected with the ploidy analyser, and adjusts fluorescence signal value to 100
Place is set as check sample；It is detected after will test sample dyeing with the ploidy analyser, records fluorescence signal value, it is included by machine
Analysis system, will test the peak value figure mean value of sample compared with check sample peak value figure, determine detection sample ploidy size；
Wherein, nucleus dye liquor is CyStain DNA1Step.
Present embodiment fixer non-volatility and inflammability long-term preservation or can be carried, be suitble in Experimental Base
Laboratory is posted go back to after sampling or detection unit uses, mainly for no the ploidy analyser or the instrument not in sampling location
Under the conditions of use.
The salmon trout embryo of 20 batches is fostered to adult fish, and carries out Ploidy Identification again in young stage, adult fish phase respectively.
Salmon trout embryonic period, embryonic phase living body ploidy identification result accuracy rate 99.9% of the present invention.The present invention is by salmon trout Ploidy detection time advance
To within 20 days, the period that more times of production of hybrid seeds conditions of salmon trout are groped and polyploid breeding is verified is substantially reduced, to accelerate salmon
Trout genetic progress has established solid foundation.
Specific embodiment 2: the difference of present embodiment and specific embodiment one is: thin in step 2 culture dish
Karyon dye liquor is 300ml, wherein 200 μ l nucleus dye liquors are placed in culture dish edge, remaining 100 μ l nucleus dye liquor is placed in training
Ware center is supported, living body embryo is placed in central 100 μ l nucleus dye liquors, and chopping rear-inclined culture dish merges nucleus dye liquor
Embryonic cell suspension is made.Present embodiment step 3 rinses culture dish with 400 μ l nucleus dye liquors.Other steps and parameter
It is identical as embodiment one.
Present embodiment step 3 is all collected all embryonic tissues into embryonic cell suspension with nucleus dye liquor.
Present embodiment single sample coloring agent handles the time as 1min, and detection speed is 2/min, and single sample reagent disappears
Consumption is only 700 μ l.
Specific embodiment 3: the difference of present embodiment and specific embodiment one or two is: detection sample is temporary
Storage must be protected from light.Other steps and parameter are identical as embodiment one or two.
Claims (4)
1. the method that a kind of pair of salmon trout embryo's living body carries out Ploidy Identification, it is characterised in that this method sequentially includes the following steps:
One, salmon trout eyespot phase embryo is individually temporarily put under the conditions of 5~8 DEG C, and keeps embryo's air exchange not anoxic；
Two, salmon trout eyespot phase embryo is pierced through with tip tweezers, releases the liquid fat in ovum, clamps embryo with tweezers later
Tire stirs in PBS buffer solution and washes excess oil, is subsequently placed in the nucleus dye liquor of culture dish, by ovum skin and living body embryo
Tire chopping shakes up embryonic cell suspension is made；
Three, it rinses culture dish with nucleus dye liquor to mix with step 2 embryonic cell suspension, then with 30 μm of aperture strainers to embryo
Fetus cells mixing suspension is filtered；It takes filtrate to be refiltered once with 30 μm of aperture strainers, obtains detection sample；
Four, the known salmon trout diploid sample of dyeing is detected with the ploidy analyser, and adjusts fluorescence signal value to setting at 100
It is set to check sample；It is detected after will test sample dyeing with the ploidy analyser, records fluorescence signal value, point carried by machine
Analysis system will test the peak value figure mean value of sample compared with check sample peak value figure, determine the ploidy size of detection sample.
2. the method that a kind of pair of salmon trout embryo's living body according to claim 1 carries out Ploidy Identification, it is characterised in that step
Nucleus dye liquor is 300ml in rapid two culture dish, wherein 200 μ l nucleus dye liquors are placed in culture dish edge, remaining 100 μ l is thin
Karyon dye liquor is placed in culture dish center, and living body embryo is placed in central 100 μ l nucleus dye liquors, and chopping rear-inclined culture dish will
Nucleus dye liquor combination system is at embryonic cell suspension.
3. the method that a kind of pair of salmon trout embryo's living body according to claim 1 carries out Ploidy Identification, it is characterised in that inspection
Originally temporarily storage must be protected from light test sample.
4. the method that a kind of pair of salmon trout embryo's living body according to claim 1 carries out Ploidy Identification, it is characterised in that thin
Karyon dye liquor is 1 Step of CyStain DNA.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810631986.0A CN109100335A (en) | 2018-06-19 | 2018-06-19 | The method that a kind of pair of salmon trout embryo's living body carries out Ploidy Identification |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810631986.0A CN109100335A (en) | 2018-06-19 | 2018-06-19 | The method that a kind of pair of salmon trout embryo's living body carries out Ploidy Identification |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109100335A true CN109100335A (en) | 2018-12-28 |
Family
ID=64796960
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810631986.0A Pending CN109100335A (en) | 2018-06-19 | 2018-06-19 | The method that a kind of pair of salmon trout embryo's living body carries out Ploidy Identification |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109100335A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN204560633U (en) * | 2015-04-30 | 2015-08-19 | 中国水产科学研究院黑龙江水产研究所 | A kind of Experimental fish class embryonic development culture apparatus |
| CN106259132A (en) * | 2016-10-17 | 2017-01-04 | 大连海洋大学 | A kind of method obtaining red fin east Androgenesis haploid |
| CN106376501A (en) * | 2016-10-24 | 2017-02-08 | 大连海洋大学 | Method for producing loach tetraploid |
| CN106957888A (en) * | 2017-04-11 | 2017-07-18 | 武汉百科金典生物科技有限公司 | A kind of method based on the ploidy analyser Rapid identification prelarva ploidy |
-
2018
- 2018-06-19 CN CN201810631986.0A patent/CN109100335A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN204560633U (en) * | 2015-04-30 | 2015-08-19 | 中国水产科学研究院黑龙江水产研究所 | A kind of Experimental fish class embryonic development culture apparatus |
| CN106259132A (en) * | 2016-10-17 | 2017-01-04 | 大连海洋大学 | A kind of method obtaining red fin east Androgenesis haploid |
| CN106376501A (en) * | 2016-10-24 | 2017-02-08 | 大连海洋大学 | Method for producing loach tetraploid |
| CN106957888A (en) * | 2017-04-11 | 2017-07-18 | 武汉百科金典生物科技有限公司 | A kind of method based on the ploidy analyser Rapid identification prelarva ploidy |
Non-Patent Citations (7)
| Title |
|---|
| 刘海金 等: "真鲷精子诱导牙鲆减数分裂雌核发育 ", 《水产学报》 * |
| 李雅娟 等: "利用天然四倍体泥鳅生产全三倍体泥鳅的初步研究 ", 《东北农业大学学报》 * |
| 杜方勇 等: "6-DMAP诱导扇贝异源三倍体的细胞学观察 ", 《水产学报》 * |
| 杨翟平 等: "虹鳟胚胎组织单细胞悬液制备方法的研究", 《水产学杂志》 * |
| 王炳谦 等: "虹鳟杂交胚胎(2n♀×3n♂)的发育状况及其DNA倍性分析 ", 《中国水产科学》 * |
| 贾光风 等: "大鳞副泥鳅(2n♀)×泥鳅(4n♂)杂种后代染色体带型及FISH分析 ", 《东北农业大学学报》 * |
| 郑春静 等: "用流式细胞仪检测大黄鱼三倍体 ", 《细胞生物学杂志》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104585091B (en) | Subfamily distant hybridization for German mirror carp and megalobrama amblycephala and application of tetraploid hybrid fishes | |
| CN104585083B (en) | Method for distant hybridization between subfamilies of cyprinus carpio and megalobrama amblycephala | |
| CN104388519B (en) | The method utilizing zebrafish embryo test natural plant extracts acute toxicity | |
| CN104704360A (en) | Gender, viability and/or developmental stage determination of avian embryos in ovo | |
| CN105115951A (en) | Method for rapidly evaluating damage effect on hepatic function of zebra fishes by compounds | |
| CN105961251B (en) | A kind of construction method of lower oxygen concentration resistance megalobrama amblycephala | |
| CN106492232B (en) | A method of with zebra fish evaluation myocardial damage inducer toxicity and therapeutic agent effect | |
| CN109220911B (en) | Application of glucose and ethanol in synergistic regulation and control of cardiovascular development of zebra fish | |
| CN106550909B (en) | The method of fancy carp and megalobrame amblycephala subfamily distant hydridization | |
| CN102893938B (en) | Subfamily distant hybridization method for Xenocypris davidi Bleeker and Erythroculter ilishaeformis Bleeker | |
| CN101302561B (en) | Method for identifying Cynoglossus semilaevis genetic sex and WW superfemale fish | |
| CN106957888A (en) | A kind of method based on the ploidy analyser Rapid identification prelarva ploidy | |
| CN107318719B (en) | Method for inducing gynogenesis of grass carp by aid of koi sperms and application of gynogenesis grass carp | |
| CN110214753A (en) | Method for building up and the application of fish products system are preced with by the method and tetraploid gold of distant hybridization breeding tetraploid Jin Guanyu | |
| CN109100188A (en) | The method that a kind of couple of salmon trout embryo carries out Ploidy Identification | |
| CN105169415B (en) | A method of screening has protection zebra fish liver function reactive compound | |
| CN109100335A (en) | The method that a kind of pair of salmon trout embryo's living body carries out Ploidy Identification | |
| CN106376501A (en) | Method for producing loach tetraploid | |
| CN104297222B (en) | Zebrafish embryo alcoholic liver detecting model and construction method and application of zebrafish embryo alcoholic liver detecting model | |
| Goodrich | A study of the development of Mendelian characters in Oryzias latipes | |
| CN103091140A (en) | Preparation method of shrimp germ cell chromosome | |
| AU2021104677A4 (en) | Method for ploidy identification of salmon and trout embryos in vivo | |
| CN112577803B (en) | Method for preparing turtle chromosome specimen by using peripheral blood cells | |
| CN108823277A (en) | The method of sturgeon Genome Size is quickly measured with Flow Cytometry | |
| CN102586392B (en) | Quality detection method for unfertilized egg of turbo |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |