CN106834473A - A kind of method of fish-egg, larva and juvenile taxonomic identification - Google Patents
A kind of method of fish-egg, larva and juvenile taxonomic identification Download PDFInfo
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- CN106834473A CN106834473A CN201710086342.3A CN201710086342A CN106834473A CN 106834473 A CN106834473 A CN 106834473A CN 201710086342 A CN201710086342 A CN 201710086342A CN 106834473 A CN106834473 A CN 106834473A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
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- C12Q2600/156—Polymorphic or mutational markers
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Abstract
The present invention relates to a kind of fish-egg, the method of larva and juvenile taxonomic identification, it is characterized in that the adult fish sample in waters is investigated in collection first, and taxonomic identification is carried out to adult fish sample, the various fish of taxonomic identification are extracted into DNA, and enter performing PCR amplification sequencing respectively to the sequences of CO I, the sequence libraries of CO I of the investigation waters fish are set up accordingly, then the fish-egg for collecting will be investigated, amplification sequencing is carried out to the sequences of CO I with same procedure after larva and juvenile sample number, sequencing result compares with the sequence libraries of CO I set up, so as to identify fish-egg, the species of larva and juvenile.The present invention can carry out quick taxonomic identification to the fish-egg of fish early development stage, larva and juvenile, overcome that conventional fish-egg, larva and juvenile identification be difficult, the deficiency that time-consuming, workload is big, it is shown that it is quick, accurately and efficiently advantage.
Description
Technical field
The present invention relates to fishery field, and in particular to a kind of fish-egg, the method for larva and juvenile taxonomic identification.
Background technology
Fish early stage resource investigation is the stock number enquiry based work carried out as object with the fish early life history stage,
It is an important means for carrying out ecology of fishes and fishery biology research.Fish early stage resource investigation can calculate that fish are numerous
Grow Population, estimate recruitment stock quantity, scientifically predict the dynamics of fish population, be fishery conservation and rationally open
Hair is utilized and provides scientific basis.During early stage resource investigation, it is necessary to gather fish-egg, larva and juvenile carry out taxonomic identification and
Quantity statistics, conventional taxonomic identification needs certain rudimentary knowledge and the worker of experience accumulation is estimated and microscope
Observation, workload is big, and elapsed time is long, and some bad assurances of Identification of Species accuracy.In fish-egg, larva and juvenile investigation sampling
When quantity is larger, conventional taxonomic identification method just shows clearly disadvantageous.
The content of the invention
The invention aims to overcome the shortcomings of conventional taxonomic identification method, there is provided a kind of fish-egg, larva and juvenile classification
The method of identification.The present invention to achieve the above object, is adopted the following technical scheme that：
A kind of method of fish-egg, larva and juvenile taxonomic identification, comprises the following steps：
(1) the adult fish sample in collection investigation waters, and taxonomic identification is carried out to adult fish sample, by the various of taxonomic identification
Fish extract DNA, and enter performing PCR amplification sequencing respectively to the sequences of CO I, and the sequence numbers of CO I of the investigation waters fish are set up accordingly
According to storehouse；
(2) fish-egg, the larva and juvenile sample number for collecting will be investigated, fish-egg, the sequences of larva and juvenile CO I are entered with same procedure
Row amplification sequencing；
(3) fish-egg, the sequence results of larva and juvenile CO I and the sequence libraries of CO I set up are compared, from
And identify the species of fish-egg, larva and juvenile.
DNA extraction method is described in step (1)：The fresh muscle of adult fish or isozyme are taken, is fixed with absolute ethyl alcohol, egg
White enzyme K digestion, phenol-imitative conventional method extracting DNA, -20 DEG C save backup.
The described sequence universal amplification primers of CO I are：Forward primer：
5-TCAACCAACCACAAAGACATTGGCAC-3, Reverse primer：
5-TAGACTTCTGGGTGGCCAAAGAATCA-3。
Described pcr amplification reaction system is：95 DEG C of predegenerations 3 minutes；Each circulation 94 DEG C 30 seconds, 56 DEG C 30 seconds,
72 DEG C 50 seconds, totally 30 circulation；Last 72 DEG C extend 6 minutes；After PCR primer is detected through 1% agarose gel electrophoresis, with purifying
Kit is reclaimed, sequencing.
The present invention only needs to be preserved with absolute ethyl alcohol after collection fish-egg, larva and juvenile, it is not necessary to complicated preservation condition, while
This is relatively conservative from the sequences of CO I fragment sets up the sequence libraries of investigation waters fish CO I, corresponding to fish-egg, larva and juvenile
Fragment carries out amplification sequencing, and the method for then being compared with the sequence libraries of CO I is entered to investigation waters fish-egg, larva and juvenile
Row taxonomic identification.The method identification degree of accuracy is high, and elapsed time is short, and workload is small, overcomes the weak point of previous methods, shows
Shown that it is quick, accurately and efficiently advantage.
Specific embodiment
The present invention is a kind of fish-egg, the method for larva and juvenile taxonomic identification, using step once：
1st, in investigation waters collection adult fish sample, and taxonomic identification is carried out to adult fish sample, takes fresh muscle or fin ray group
Knit, fixed with absolute ethyl alcohol, protease K digesting, phenol-imitative conventional method extracting DNA, -20 DEG C save backup；
2nd, the relatively conservative sequence fragments of CO I of selection, be from universal amplification primer：Forward primer：5-
TCAACCAACCACAAAGACATTGGCAC-3, Reverse primer：
5-TAGACTTCTGGGTGGCCAAAGAATCA-3；
3rd, pcr amplification reaction system is：95 DEG C of predegenerations 3 minutes；Each circulation 94 DEG C 30 seconds, 56 DEG C 30 seconds, 72 DEG C
50 seconds, totally 30 circulations；Last 72 DEG C extend 6 minutes；
4th, after PCR primer is detected through 1% agarose gel electrophoresis, reclaimed with purification kit, sequencing；
5th, the sequences of CO I of various fish are collected, sets up the sequence libraries of CO I；
6th, in the collection of investigation waters fish-egg, larva and juvenile sample are fixed with absolute ethyl alcohol, and a part of sample is selected at random simultaneously
Numbering, according to step 1 methods described extract DNA and be stored in -20 DEG C it is standby；
7th, according to experimental arrangement described in step 2-4, sequencing obtains each fish-egg, the sequences of CO I of larva and juvenile sample；
8th, acquired each fish-egg, the sequences of CO I of larva and juvenile sample and the sequence libraries of CO I set up are compared
Analysis, taxonomic identification is carried out to fish-egg, larva and juvenile sample.
Claims (4)
1. a kind of fish-egg, the method for larva and juvenile taxonomic identification, comprise the following steps：
(1) the adult fish sample in collection investigation waters, and taxonomic identification is carried out to adult fish sample, by the various fish of taxonomic identification
DNA is extracted, and enters performing PCR amplification sequencing respectively to the sequences of CO I, the sequence libraries of CO I of the investigation waters fish are set up accordingly；
(2) fish-egg, the larva and juvenile sample number for collecting will be investigated, fish-egg, the sequences of larva and juvenile CO I are expanded with same procedure
Increase sequencing；
(3) fish-egg, the sequence results of larva and juvenile CO I and the sequence libraries of CO I set up are compared, so as to reflect
Determine the species of fish-egg, larva and juvenile.
2. a kind of fish-egg according to claim 1, the method for larva and juvenile taxonomic identification, it is characterised in that：Institute in step (1)
Stating DNA extraction method is：The fresh muscle of adult fish or isozyme are taken, is fixed with absolute ethyl alcohol, protease K digesting, phenol-imitative conventional
Method extracts DNA, and -20 DEG C save backup.
3. a kind of fish-egg according to claim 1, the method for larva and juvenile taxonomic identification, it is characterised in that：The sequences of CO I are general
Amplimer is：Forward primer：5-TCAACCAACCACAAAGACATTGGCAC-3, Reverse primer：5-
TAGACTTCTGGGTGGCCAAAGAATCA-3。
4. a kind of fish-egg according to claim 1, the method for larva and juvenile taxonomic identification, it is characterised in that：Described PCR expands
Increasing reaction system is：95 DEG C of predegenerations 3 minutes；Each circulation 94 DEG C 30 seconds, 56 DEG C 30 seconds, 72 DEG C 50 seconds, totally 30 circulation；
Last 72 DEG C extend 6 minutes；After PCR primer is detected through 1% agarose gel electrophoresis, reclaimed with purification kit, sequencing.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710086342.3A CN106834473A (en) | 2017-02-17 | 2017-02-17 | A kind of method of fish-egg, larva and juvenile taxonomic identification |
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| CN201710086342.3A CN106834473A (en) | 2017-02-17 | 2017-02-17 | A kind of method of fish-egg, larva and juvenile taxonomic identification |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108651337A (en) * | 2018-05-15 | 2018-10-16 | 江西省水产科学研究所 | A kind of production Drifting egg fish early stage different developmental phases form microscopy observation method |
| CN109055568A (en) * | 2018-07-25 | 2018-12-21 | 江汉大学 | A kind of emarginated tail intends Chang DNA bar code sequence and its application |
| CN109055515A (en) * | 2018-08-28 | 2018-12-21 | 浙江海洋大学 | A kind of molecular biology method identifying Xinjiang Eerqisihe River north palpus loach cryptic species |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102229997A (en) * | 2011-06-01 | 2011-11-02 | 中国水产科学研究院淡水渔业研究中心 | Method for rapidly classifying and identifying fish larvae |
| CN102533997A (en) * | 2012-01-04 | 2012-07-04 | 中国科学院水生生物研究所 | Method for identifying fish early individual by combining molecular biology and morphology |
| CN105349645A (en) * | 2015-11-10 | 2016-02-24 | 中国水产科学研究院淡水渔业研究中心 | Roe and fry quick classification and determination methods based on CoI DNA bar codes |
| CN106282380A (en) * | 2016-09-30 | 2017-01-04 | 中国长江三峡集团公司中华鲟研究所 | A kind of Fish based on DNA bar code technology and raw meat product quality monitoring and managing method thereof and system thereof |
-
2017
- 2017-02-17 CN CN201710086342.3A patent/CN106834473A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102229997A (en) * | 2011-06-01 | 2011-11-02 | 中国水产科学研究院淡水渔业研究中心 | Method for rapidly classifying and identifying fish larvae |
| CN102533997A (en) * | 2012-01-04 | 2012-07-04 | 中国科学院水生生物研究所 | Method for identifying fish early individual by combining molecular biology and morphology |
| CN105349645A (en) * | 2015-11-10 | 2016-02-24 | 中国水产科学研究院淡水渔业研究中心 | Roe and fry quick classification and determination methods based on CoI DNA bar codes |
| CN106282380A (en) * | 2016-09-30 | 2017-01-04 | 中国长江三峡集团公司中华鲟研究所 | A kind of Fish based on DNA bar code technology and raw meat product quality monitoring and managing method thereof and system thereof |
Non-Patent Citations (1)
| Title |
|---|
| 曲钦岳等: "《当代百科知识大词典》", 30 June 1989 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108651337A (en) * | 2018-05-15 | 2018-10-16 | 江西省水产科学研究所 | A kind of production Drifting egg fish early stage different developmental phases form microscopy observation method |
| CN109055568A (en) * | 2018-07-25 | 2018-12-21 | 江汉大学 | A kind of emarginated tail intends Chang DNA bar code sequence and its application |
| CN109055515A (en) * | 2018-08-28 | 2018-12-21 | 浙江海洋大学 | A kind of molecular biology method identifying Xinjiang Eerqisihe River north palpus loach cryptic species |
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Application publication date: 20170613 |