CN112831573A - SNP primer and detection method for identifying sturgeon germplasm - Google Patents
SNP primer and detection method for identifying sturgeon germplasm Download PDFInfo
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Abstract
The invention discloses an SNP primer and a detection method for identifying sturgeon germplasm. The invention develops the germplasm specificity SNP locus by comparing acipenser schrencki, acipenser siberius and huso dauricus mitochondrial genomes, invents 2 pairs of peripheral primers and 7 middle primers, can carry out germplasm identification on acipenser schrencki, acipenser sibericus and huso dauricus, and is applied to the identification of sturgeon germplasm resources. The SNP primer is suitable for the SNaPshot detection technology, has the characteristics of high efficiency, accuracy and low cost, can accurately detect the mutation site, and can meet the requirement of high-flux detection.
Description
Technical Field
The invention belongs to the technical field of sturgeon identification, and particularly relates to an SNP primer and a detection method for sturgeon germplasm identification.
Background
Sturgeon is one of the earliest vertebrates of existing origin. Sturgeons belong to the subclasses Osteichthyes, Apyratiales, Chordariales order and Acipenseridae order. Sturgeons are generally known as fish of the order sturgeon. Sturgeon fish is a kind of fish with polyploid origin, has a large number of chromosomes, and has various natural hybrid species, and the hybrid species of sturgeon fish has high fertility biological characteristics. Sturgeons, particularly hybrid sturgeons, are difficult to morphologically distinguish, have strong morphological subjectivity, and are easy to make misjudgments. The fish germplasm molecular identification technology is a new technology developed by utilizing molecular biology technology and aiming at researching biological species identification and phylogenetic relationship, and is used for identifying varieties by identifying differences on DNA level. A single nucleotide polymorphism is a sequence polymorphism caused by a variation of a single nucleotide at a specific site in a genomic DNA sequence. The SNP spreads over the whole genome, and one SNP is found at 1900bp on average in human beings, so that the SNP has higher genetic stability compared with a microsatellite, can realize detection automation, and is very suitable for high-speed detection of germplasm identification. Sequencing is the most accurate genotyping technology, the most intuitive, efficient and pollution-free mutation detection technology, but the cost is high. The SNaPshot detection technology adopted in the research is also called micro sequencing, has the characteristics of high efficiency, accuracy and low cost, can accurately detect mutation sites, and can meet the requirement of high-flux detection.
Disclosure of Invention
The invention aims to provide an SNP primer and a detection method for identifying sturgeon germplasm, which solve the existing problems: aiming at the technical defects in the sturgeon germplasm identification, the intuitive, accurate and efficient germplasm identification method for the Siberian sturgeon, the Acipenser schrenki and the huso dauricus is provided. And provides the application of the SNP primer in identifying sturgeon germplasm.
In order to solve the technical problems, the invention is realized by the following technical scheme:
the invention relates to an SNP primer and a detection method for identifying sturgeon germplasm, wherein a primer combination of AF + AR + SNP1/SNP2/SNP3/SNP4 or BF + BR + SNP5/SNP6/SNP7 is adopted to identify the sturgeon germplasm. The detection method comprises the steps of sample collection and DNA extraction, primer amplification of SNP sites and SNaPshot detection of SNP sites, wherein the sample collection and the genome DNA extraction comprise the following steps:
the method comprises the following steps: collecting a tail fin sample of the sturgeon to be detected;
step two: fixing the tail fin sample in absolute ethyl alcohol, and storing at 4 ℃ for later use.
Further, the SNP primer for identifying the germplasm of the sturgeon comprises 2 pairs of peripheral primers and 7 intermediate primer sets, wherein the nucleotide sequence of each primer is shown in the following table:
further, the method for detecting the SNP site by the SNaPshot comprises the following steps:
the method comprises the following steps: and (3) peripheral primer amplification target fragment conditions: the total volume of the PCR reaction was 50. mu.L, where: template DNA 100ng, I-5TMHigh-Fidelity Master Mix (MCLAB) 25. mu.L, peripheral primers (10. mu. mol/L) 2. mu.L each, supplemented with sterile double distilled water to 50. mu.L;
step two: peripheral primer amplification of target fragment program: 2min at 98 ℃; 30 cycles of 98 ℃ for 10s, 57 ℃ for 10s, and 72 ℃ for 10 s; extension at 72 ℃ for 5 min.
Step three: digesting the PCR product by SAP enzyme to remove the residual dNTP and primer in the PCR system, wherein the reaction conditions are as follows: 1h at 37 ℃, 15min at 75 ℃ and infinity at 4 ℃;
step four: amplifying SNP sites by using the intermediate primer, extending single strands:
reaction system: 5 μ L
Template 2. mu.L after SAP digestion
ABI SnapShot multiplex Mix 2μL
1 mu L of intermediate primer;
step five: reaction conditions are as follows:
the SNP primers are applied to obtain the corresponding genotypes of 30 individuals of Siberian sturgeon, Acipenser schrenki and huso dauricus, and the genotypes are shown in the following table:
the invention is proved by experiments that: primer combinations of AF + AR + SNP1/SNP2/SNP3/SNP4 or BF + BR + SNP5/SNP5/SNP7 can be used for identifying the germplasm of sturgeons.
The invention has the following beneficial effects:
1. according to the invention, 2 pairs of peripheral primers and 7 pieces of intermediate primers are designed according to the developed idioplasmic SNP loci of the Acipenser schrenki, the Acipenser sibiricus and the huso dauricus, and the idioplasm identification of the Acipenser schrenki, the Acipenser sibiricus and the huso dauricus can be carried out through the matching of the peripheral primers and the intermediate primers, so that the invention can be applied to the idioplasm resource identification of the Acipenser schrenki.
2. The SNP primer is suitable for the SNaPshot detection technology, has the characteristics of high efficiency, accuracy and low cost, can accurately detect the mutation site, and can meet the requirement of high-flux detection.
Of course, it is not necessary for any product in which the invention is practiced to achieve all of the above-described advantages at the same time.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a table of peripheral primer information according to the present invention;
FIG. 2 is a table of the middle primer information in the present invention;
FIG. 3 shows the germplasm-specific SNP sites of Acipenser sibirica, Acipenser schrenki, huso dauricus according to the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1-3, the present invention relates to an SNP primer and detection method for identifying sturgeon germplasm, including sample collection and genomic DNA extraction, primer design and SNaPshot detection of SNP sites. The sample collection and genomic DNA extraction comprises the following steps:
the method comprises the following steps: respectively collecting tail fin samples of 30 tails of Siberian sturgeon parent fish, 30 tails of Acipenser schrenki parent fish and 30 tails of huso dauricus parent fish;
step two: fixing the tail fin sample in absolute ethyl alcohol, and storing at 4 ℃ for later use.
Step three: a sample of about 50mg of sturgeon fin-shaped strip is cut, liquid nitrogen is ground into powder, a magnetic bead method genome DNA (animal) extraction kit (Shanghai Biotechnology engineering Co., Ltd.) is adopted for DNA extraction, and the quality and concentration of DNA are measured by agarose gel electrophoresis and a NanoDrop lite (Thermo Scientific) ultraviolet spectrophotometer.
The primer design comprises the following steps:
the mitochondrial genome sequences of Acipenser sibirica, Acipenser schrenki and huso dauricus are obtained from GenBank databases, and the GenBank accession numbers are JQ045341.1, KC820796.1 and KJ402277.1 respectively. The MEGA 6.0 software is used for genome sequence comparison, two sequence fragments of 46-849bp and 1725-2383bp are selected according to the comparison result, 10 SNP sites are contained, and the Primer premier5 software is used for designing peripheral Primer amplification gene fragments, wherein the peripheral Primer information is shown in Table 1. SNP sites are detected by adopting a SNaPshot detection technology, so that 10 intermediate primers are designed for detecting the SNP sites, and the information of the intermediate primers is shown in Table 2.
TABLE 1 peripheral primer information
TABLE 2 intermediate primer information
| Primer name | Primer sequences | Direction of primer extension |
| SNP1 | TTTTTTTTTAAGCATAACACTGAAGATGTTAAGATG | Forward direction |
| SNP2 | TTTTTTTTTTTTTTTTTTTTTTTAACTCTGGCTGAGTCGAGCTT | Reverse direction |
| SNP3 | TTTTTTTTTTTTTTTTTTTTTTTTTTGCGTAAAGCGTGATTAAAGGAT | Forward direction |
| SNP4 | TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTCAACCACGAAGGTAGCTCTACC | Forward direction |
| SNP5 | TTTTTTTTTTTGATTGTTTTATCGGGATAATAGGGT | Reverse direction |
| SNP6 | TTTTTTTTTTTTTTTTTTTAGGGTCTTCTCGTCTTATGGG | Reverse direction |
| SNP7 | TTTTTTTTTTTTTTTTTTTTTTTTTTCGACCACGGAGGACAAAA | Forward direction |
| SNP8 | TTTTTTTTTTTTTTTTTTTTGGGATTAAGGGCATTCTCAC | Reverse direction |
| SNP9 | TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTACGGCGGTATATAGGCGG | Reverse direction |
| SNP10 | TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTACTGCTAAATCCACCTTCAA | Reverse direction |
The method for detecting the SNP locus by the SNaPshot comprises the following steps:
the method comprises the following steps: and (3) peripheral primer amplification target fragment conditions: the total volume of the PCR reaction was 50. mu.L, where: template DNA 100ng, I-5TMHigh-Fidelity Master Mix (MCLAB) 25. mu.L, peripheral primers (10. mu. mol/L) 2. mu.L each, supplemented with sterile double distilled water to 50. mu.L;
step two: peripheral primer amplification of target fragment program: 2min at 98 ℃; 30 cycles of 98 ℃ for 10s, 57 ℃ for 10s, and 72 ℃ for 10 s; extension at 72 ℃ for 5 min. Detecting the PCR amplification product by agarose gel electrophoresis, sequencing by Pongonidae organism company, and verifying the authenticity of the mutation site;
step three: digesting the PCR product by SAP enzyme to remove the residual dNTP and primer in the PCR system, wherein the reaction conditions are as follows: 1h at 37 ℃, 15min at 75 ℃ and infinity at 4 ℃;
step four: amplifying SNP sites by using the intermediate primer, extending single strands:
reaction system: 5 μ L
Template 2. mu.L after SAP digestion
ABI SnapShot multiplex Mix 2μL
SNP Primer 1μL；
Step five: reaction conditions are as follows:
snapshot assay
Step five: the product obtained after single-strand extension was mixed with HIDI to which an internal standard was added.
Plate mixing conditions: template 1. mu.L; HIDI 10. mu.L.
Mixing the plates, performing thermal denaturation at 95 deg.C for 5min, cooling in a refrigerator at-20 deg.C for 2-3min, and loading onto a machine.
Step six: the raw data analysis was performed with Genemapper4.0 software after 50min on the computer Applied Biosystems 3730 XL.
The results show that: primer combinations of AF + AR + SNP1/SNP2/SNP3/SNP4 or BF + BR + SNP5/SNP5/SNP7 can be used for identifying the germplasm of sturgeons.
One specific application of this embodiment is: collecting tail fin samples of 20 sturgeons of unknown species, and extracting DNA. And (3) carrying out germplasm identification on the sturgeon of unknown species by adopting two primer combinations of AF + AR + SNP1/SNP2 and BF + BR + SNP 6.
And (3) peripheral primer amplification target fragment conditions: the total volume of the PCR reaction was 50. mu.L, where: template DNA 100ng, I-5TMHigh-Fidelity Master Mix (MCLAB) 25. mu.L, peripheral primers (10. mu. mol/L) 2. mu.L each, sterile double distilled water supplemented to 50. mu.L, peripheral primers amplification of fragment of interest program: 2min at 98 ℃;30 cycles of 98 ℃ for 10s, 57 ℃ for 10s, and 72 ℃ for 10 s; extension at 72 ℃ for 5 min.
Digesting the PCR product by SAP enzyme to remove the residual dNTP and primer in the PCR system, wherein the reaction conditions are as follows: 1h at 37 ℃, 15min at 75 ℃, infinity at 4 ℃, reaction system: 5 μ L, Template 2 μ L after SAP digestion, ABI SnapShot multiplex Mix2 μ L, middle primer 1 μ L; the reaction conditions are 96 ℃ for 10s, 50 ℃ for 5s, 60 ℃ for 30s and 4 ℃ for infinity.
SNaPshot assay: the product obtained after single strand extension was mixed with HIDI with an internal standard and loaded onto an Applied Biosystems 3730XL for 50min before analysis of the raw data using Genemapper4.0 software. Referring to FIG. 3, the SNP sites and germplasm identification results of 20 sturgeons of unknown species are shown in Table 3:
TABLE 320 species and quality identification results of Acipenser sinensis
In the description herein, references to the description of "one embodiment," "an example," "a specific example" or the like are intended to mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The preferred embodiments of the invention disclosed above are intended to be illustrative only. The preferred embodiments are not intended to be exhaustive or to limit the invention to the precise embodiments disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best utilize the invention. The invention is limited only by the claims and their full scope and equivalents.
Claims (2)
1. An SNP primer for identifying sturgeon germplasm, which is characterized by comprising the following components in percentage by weight: consists of 2 pairs of peripheral primers and 7 middle primers, and the nucleotide sequences of the primers are as follows:
TABLE 1 primer information
2. The SNP primer and the detection method for identifying the germplasm of sturgeons according to claim 1, wherein the SNP primer and the detection method are characterized in that: the method for detecting the SNP locus by the SNaPshot comprises the following steps:
the method comprises the following steps: and (3) peripheral primer amplification target fragment conditions: the total volume of the PCR reaction was 50. mu.L, where: template DNA 100ng, I-5TMHigh-Fidelity Master Mix (MCLAB) 25. mu.L, peripheral primers (10. mu. mol/L) 2. mu.L each, supplemented with sterile double distilled water to 50. mu.L;
step two: peripheral primer amplification of target fragment program: 2min at 98 ℃; 30 cycles of 98 ℃ for 10s, 57 ℃ for 10s, and 72 ℃ for 10 s; extending for 5min at 72 ℃;
step three: digesting the PCR product by SAP enzyme to remove the residual dNTP and primer in the PCR system, wherein the reaction conditions are as follows: 1h at 37 ℃, 15min at 75 ℃ and infinity at 4 ℃;
step four: amplifying SNP sites by using the intermediate primer, extending single strands:
reaction system: 5 μ L
Template 2. mu.L after SAP digestion
ABI SnapShot multiplex Mix 2μL
1 mu L of intermediate primer;
step five: reaction conditions are as follows:
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114107521A (en) * | 2021-12-20 | 2022-03-01 | 中国海关科学技术研究中心 | Primer probe composition and kit for detecting pure Siberian sturgeon nuclear gene and application of primer probe composition and kit |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105002170A (en) * | 2015-07-31 | 2015-10-28 | 中国长江三峡集团公司 | Amur sturgeon germplasm molecular marker identification kit and application thereof |
| CN105063192A (en) * | 2015-07-31 | 2015-11-18 | 中国长江三峡集团公司 | Molecular marker for identifying amur sturgeon germplasm and application of molecular marker |
| EP3199643A1 (en) * | 2015-12-31 | 2017-08-02 | Ustav biologie obratlovcu AV CR, v.v.l. | Method of identification of european freshwater fish and hybrides in biological materials by s7icaps method |
| CN108660220A (en) * | 2018-05-21 | 2018-10-16 | 中国水产科学研究院 | One group of sturgeon SNP marker, sturgeon production of hybrid seeds appraisal procedure and genetic evaluation method |
| CN110747279A (en) * | 2019-11-21 | 2020-02-04 | 南京师范大学 | Fugu obscurus SNP molecular marker and application thereof in genetic breeding |
| CN110923336A (en) * | 2019-12-27 | 2020-03-27 | 四川省农业科学院水产研究所(四川省水产研究所) | Primer, molecular marker and method for identifying germplasm of Acipenser dabryanus and Acipenser sinensis |
-
2021
- 2021-03-22 CN CN202110301543.7A patent/CN112831573B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105002170A (en) * | 2015-07-31 | 2015-10-28 | 中国长江三峡集团公司 | Amur sturgeon germplasm molecular marker identification kit and application thereof |
| CN105063192A (en) * | 2015-07-31 | 2015-11-18 | 中国长江三峡集团公司 | Molecular marker for identifying amur sturgeon germplasm and application of molecular marker |
| EP3199643A1 (en) * | 2015-12-31 | 2017-08-02 | Ustav biologie obratlovcu AV CR, v.v.l. | Method of identification of european freshwater fish and hybrides in biological materials by s7icaps method |
| CN108660220A (en) * | 2018-05-21 | 2018-10-16 | 中国水产科学研究院 | One group of sturgeon SNP marker, sturgeon production of hybrid seeds appraisal procedure and genetic evaluation method |
| CN110747279A (en) * | 2019-11-21 | 2020-02-04 | 南京师范大学 | Fugu obscurus SNP molecular marker and application thereof in genetic breeding |
| CN110923336A (en) * | 2019-12-27 | 2020-03-27 | 四川省农业科学院水产研究所(四川省水产研究所) | Primer, molecular marker and method for identifying germplasm of Acipenser dabryanus and Acipenser sinensis |
Non-Patent Citations (6)
| Title |
|---|
| R OGDEN等: "Sturgeon conservation genomics: SNP discovery and validation using RAD sequencing", MOL ECOL * |
| RUI RUAN等: ""Screening and identification of female-specific DNA sequences in octaploid sturgeon using comparative genomics with high-throughput sequencing"", 《GENOMICS》 * |
| 孔杰等: "" 施氏鲟、西伯利亚鲟、达氏鳇及其杂交种种质鉴定方法的建立及其应用"", 《中国水产科学》 * |
| 孔杰等: ""分子标记在鲟鱼研究中的应用"", 《农技服务》 * |
| 牛晓锋等: ""鲟鱼分子鉴定方法的整合应用"", 《生物多样性》 * |
| 董颖等: "鲟鱼种质鉴定技术的发展及应用", 中国水产 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114107521A (en) * | 2021-12-20 | 2022-03-01 | 中国海关科学技术研究中心 | Primer probe composition and kit for detecting pure Siberian sturgeon nuclear gene and application of primer probe composition and kit |
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