CN115896298A - Siniperca chuatsi X chromosome molecular marker primer and application thereof - Google Patents
Siniperca chuatsi X chromosome molecular marker primer and application thereof Download PDFInfo
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Abstract
The invention discloses a molecular marker primer of siniperca chuatsi X chromosome, which is characterized by comprising the following components in parts by weight: the primer is a primer pair 1 or a primer pair 2, the primer pair 1 comprises a Contig-1 upstream primer and a Contig-1 downstream primer, and the primer pair 2 comprises a Contig-2 upstream primer and a Contig-2 downstream primer, wherein the nucleotide sequence of the Contig-1 upstream primer is shown in SEQ ID NO:1, the nucleotide sequence of the Contig-1 downstream primer is shown as SEQ ID NO:2, the nucleotide sequence of the Contig-2 upstream primer is shown as SEQ ID NO:3, the nucleotide sequence of the Contig-2 downstream primer is shown as SEQ ID NO:4, respectively. Also discloses a method for identifying the YY super-male mandarin fish and application of the primer or the method in the full male breeding of the mandarin fish.
Description
Technical Field
The invention belongs to the technical field of fish sex chromosome composition identification, and particularly relates to a siniperca chuatsi X chromosome molecular marker primer and application thereof.
Background
In recent years, with the rapid development of molecular biology technology, various sex chromosome specific markers of fish have been developed, such as nile tilapia (Oreochromis niloticus), rainbow trout (Oncorhynchus mykiss), pelteobagrus fulvidraco (pelteobagrus fulvidraco), cynoglossus semilaevis (cynoglossus semilaevis), and takifugurus rubrus (takifugurus), but these markers are mainly obtained by molecular marker development technologies such as microsatellite (SSR), restriction site associated DNA (RAD), and amplified fragment length polymorphism (amplified fragment length polymorphism, AFLP), and these molecular marker development methods are long in time consumption, large in workload, and low in success rate. The appearance of high-throughput sequencing technology provides a new high-efficiency means for developing molecular markers, and specific molecular markers of sex chromosomes of snakeheads (Channa argus) and large yellow croakers (larmichtysoscea) are successfully developed by a high-throughput sequencing method at present.
Siniperca chuatsi (Sinipercachuatsi) belongs to Perciformes, sinipercidae and Sinipercica, and is commonly called Sinipercica and Mandarin Fish, and the English names thereof mean Chinese bass (Chinese perch) and Mandarin Fish (Mandarin Fish). The siniperca chuatsi is a unique and rare fish in fresh water in China, has delicious meat, rich nutrition and no muscle thorns, and is deeply favored by consumers at home and abroad. In 2-5 months every year, the demand of the mandarin fish in China is large, the sale price is high, but the storage amount of the mandarin fish in the pond is small. In recent years, farmers choose to sell mandarin fish after wintering period in order to obtain higher profit. The growth of siniperca chuatsi has gender polymorphism, during 2-9 growth months before the overwintering period, the growth speed of female fish is faster than that of male fish, and during 9-14 growth months after the overwintering period, the female fish reaches sexual maturity and the growth speed is slowed down, and at the moment, the growth speed of the male fish is faster than that of the female fish. Therefore, the method has extremely high economic value for cultivating the all-male siniperca chuatsi.
In the past karyotype analysis of siniperca chuatsi, no obvious sex chromosome is found, but through sequencing of a male and female genome and development of a siniperca chuatsi male specific molecular marker, a Y chromosome specific DNA fragment exists in a male fish genome, so that the sex of the mandarin fish is judged to be XX/XY type, namely male heterozygosis. The parthenocarpy breeding of the mandarin fish needs to obtain YY super-male fish firstly, and the main way of obtaining the super-male fish is to mate pseudo female fish obtained by estrogen induction with normal male fish so as to obtain YY super-male fish offspring, wherein the super-male fish accounts for 25% of the offspring theoretically. And secondly, mating the obtained super-male fish with normal female fish to obtain male fish offspring with the genotype of XY totally. The traditional method for screening XY male fish and YY super male fish is to judge the sex chromosome composition of male parent through test cross experiment. The sex of the test cross offspring is verified by the mandarin fish male specificity marker, a large amount of experiment cost is consumed, and the independent breeding of the test cross parents needs to occupy more breeding resources, so the test cross method is time-consuming, labor-consuming and resource-consuming. So far, no report that the Siniperca chuatsi X chromosome specific marker can be successfully identified exists at home and abroad, so that the development of the Siniperca chuatsi X chromosome molecular marker for rapidly screening YY super-male fish has great production and application values for the full-male breeding of Siniperca chuatsi.
Disclosure of Invention
The first purpose of the invention is to provide a siniperca chuatsi X chromosome molecular marker primer, which can identify YY supermale siniperca chuatsi and XY male siniperca chuatsi on a molecular level.
The second purpose of the invention is to provide a method for identifying the YY super-male mandarin fish with the tilting mouth, which has short time consumption and accurate detection result.
The third purpose of the invention is to provide the application of the primer or the method in the full-male breeding of the siniperca chuatsi.
In order to achieve the first purpose, the invention adopts the following technical scheme:
a Siniperca Chuatsi X chromosome molecular marker primer is a primer pair 1 or a primer pair 2, wherein the primer pair 1 comprises a Contig-1 upstream primer and a Contig-1 downstream primer, and the primer pair 2 comprises a Contig-2 upstream primer and a Contig-2 downstream primer, wherein the nucleotide sequence of the Contig-1 upstream primer is shown in SEQ ID NO:1, the nucleotide sequence of the Contig-1 downstream primer is shown as SEQ ID NO:2, the nucleotide sequence of the Contig-2 upstream primer is shown as SEQ ID NO:3, the nucleotide sequence of the Contig-2 downstream primer is shown as SEQ ID NO:4, respectively.
Specifically, the nucleotide sequence of each primer from 5 'to 3' is as follows:
contig-1 upstream primer: 5'-ATTCCCATCAACCTGT-3';
contig-1 downstream primer: 5'-CACAATCTGTGCCTAC-3';
contig-2 upstream primer: 5'-TTGCGTTTAGTGTTGATTGT-3';
contig-2 downstream primer: 5'-AGACCTGCACGTCCTAA-3'.
In order to achieve the second object, the invention adopts the following technical scheme:
a method for identifying YY super-male mandarin fish comprises the following steps:
(1) Preparing a siniperca chuatsi DNA sample;
(2) Carrying out PCR amplification by using the primer pair 1 or the primer pair 2 in the claim 1 by using the DNA sample in the step (1) as a template, carrying out electrophoresis detection after the amplification reaction is finished, wherein if the electrophoresis result shows that an amplification product has an X chromosome specific strip, the detected siniperca chuatsi DNA sample is XX female siniperca chuatsi or XY male siniperca chuatsi, and if the electrophoresis result shows that the amplification product does not have the X chromosome specific strip, the detected siniperca chuatsi DNA sample is YY super male siniperca chuatsi.
In the method for identifying the YY super-male mandarin fish by tilting the mouth:
preferably, in the step (2), the DNA sample in the step (1) is used as a template, a primer pair 1 is adopted for PCR amplification, electrophoresis detection is carried out after the amplification reaction is finished, if the electrophoresis result shows that an amplification product has an X chromosome specific band of about 935bp, the detected Siniperca chuatsi DNA sample is XX female Siniperca chuatsi or XY male Siniperca chuatsi, and if the electrophoresis result shows that the amplification product does not have the X chromosome specific band of about 935bp, the detected Siniperca chuatsi DNA sample is YY supermale siniperca chuatsi.
Preferably, in the step (2), the DNA sample in the step (1) is used as a template, a primer pair 2 is adopted for PCR amplification, electrophoresis detection is carried out after the amplification reaction is finished, if the electrophoresis result shows that an amplification product has an X chromosome specific band of about 850bp, the detected Siniperca chuatsi DNA sample is XX female Siniperca chuatsi or XY male Siniperca chuatsi, and if the electrophoresis result shows that the amplification product does not have the X chromosome specific band of about 850bp, the detected Siniperca chuatsi DNA sample is YY supermale Siniperca chuatsi.
Preferably, the DNA sample preparation in the step (1) adopts a column type centrifugation method to extract the DNA of the siniperca chuatsi.
Preferably, in the step (2), a 20 μ L PCR reaction system is adopted during PCR amplification, the 20 μ L PCR reaction system comprises 50ng of DNA, 0.8 μ L of each of the Contig-1 upstream primer and the Contig-1 downstream primer of the primer pair 1 or 0.8 μ L of each of the Contig-2 upstream primer and the Contig-2 downstream primer of the primer pair 2, and 2 XTaq MasterMix 10 μ L, and ddH is used for the rest 2 And (4) supplementing oxygen.
Preferably, when the PCR is performed in the step (2), the PCR reaction procedure is as follows: firstly, pre-denaturation is carried out for 2min at 94 ℃; then carrying out denaturation at 94 ℃ for 30s, annealing at 56 ℃ for 30s and extension at 72 ℃ for 30s for 35 cycles; and final extension at 72 ℃ for 5min.
Preferably, in the step (2), agarose gel electrophoresis with a mass percentage of 1% is used for detecting the length of the amplified fragment.
In order to achieve the third object, the invention adopts the following technical scheme:
the primer or the method is applied to the full male breeding of the siniperca chuatsi.
The invention has the following beneficial effects:
(1) The method is based on the genome high-throughput sequencing data of YY supermale and XX female siniperca chuatsi, and utilizes analysis methods such as genome comparison and assembly to quickly and accurately screen out siniperca chuatsi X chromosome molecular markers, and molecular marker primers for identifying the siniperca chuatsi X chromosome are designed for the first time, and can quickly and accurately identify YY supermale fish of the siniperca chuatsi;
(2) The method establishes a method for identifying the Y super-male mandarin fish by using the Siniperca chuatsi X chromosome molecular marker primer, and can identify the YY super-male mandarin fish by completing a PCR reaction by using two pairs of specific primers.
Drawings
FIG. 1 is a flow chart of the selection and breeding of YY super-male fish in example 1;
FIG. 2 is the electrophoresis chart of YY fish offspring screened for sequencing in example 1, wherein a-f are YY test cross offspring, and g are XY test cross offspring;
FIG. 3 is a schematic diagram showing the acquisition of X chromosome-specific candidate fragments in example 1;
FIG. 4 is an electrophoresis chart of PCR products of primers Contig-1 and Contig-2 of example 1, which detected fragments with amplification sizes of 935bp and 850bp in XX female and XY male samples, respectively, and no corresponding amplification in YY super-male fish;
FIG. 5 is an electrophoretogram of PCR products of the amplification of non-X chromosome-specific fragments in XX females, XY males, and YY supermans with the remaining four primer pairs in example 1;
FIG. 6 is an electrophoretogram of PCR products of primer Contig-1 in 8 XX female, 8 XY male and 6 YY supermale samples in example 2;
FIG. 7 is an electrophoretogram of PCR products of primer Contig-2 in 8 XX female, 8 XY male and 6 YY supermale samples in example 2.
Detailed Description
The technical solutions of the present invention are described in detail below with reference to specific examples so that those skilled in the art can better understand and implement the technical solutions of the present invention. Reagents or materials used in the examples were commercially available, unless otherwise specified.
Example 1
The embodiment provides an X chromosome molecular marker primer of siniperca chuatsi, which comprises a primer pair 1 or a primer pair 2, wherein the primer pair 1 comprises a Contig-1 upstream primer and a Contig-1 downstream primer, and the primer pair 2 comprises a Contig-2 upstream primer and a Contig-2 downstream primer, wherein the nucleotide sequence of the Contig-1 upstream primer is shown in SEQ ID NO:1, the nucleotide sequence of the Contig-1 downstream primer is shown as SEQ ID NO:2, the nucleotide sequence of the Contig-2 upstream primer is shown as SEQ ID NO:3, the nucleotide sequence of the Contig-2 downstream primer is shown as SEQ ID NO:4, respectively.
Specifically, the nucleotide sequence of each primer from 5 'to 3' is as follows:
contig-1 upstream primer: 5'-ATTCCCATCAACCTGT-3';
contig-1 downstream primer: 5'-CACAATCTGTGCCTAC-3';
contig-2 upstream primer: 5'-TTGCGTTTAGTGTTGATTGT-3';
contig-2 downstream primer: 5'-AGACCTGCACGTCCTAA-3'.
The molecular marker primer of the Siniperca chuatsi X chromosome is obtained by the following steps:
(I) sequencing library construction and re-sequencing
1. Preparing a mandarin fish DNA sample:
during the propagation period, 3-tailed super-male tail fins of the mandarin fish are cut, and the super-male fish is obtained by screening through a test cross method (figure 1 and figure 2). Numbering the super-male fish SM1-SM3 in sequence, preparing a DNA sample by adopting a conventional column type centrifugation method (a general column type genome extraction kit, beijing kang is century biotechnology limited) according to the operation steps of the specification, and detecting the quality and the concentration by using 1% agarose gel electrophoresis and a micro spectrophotometer (NanoDrop 2000) to meet the requirement of high-throughput sequencing.
2. Sequencing library construction and high throughput sequencing:
taking the DNA samples of the super-male fish SM1-SM3, respectively constructing a 350-500bp fragment sequencing library, and carrying out double-End (Pair-End) PE150 sequencing by using an Illumina Hiseq X Ten platform to obtain super-male fish genome sequencing clean data:33,810,178,500bp.
The Library construction procedure was specifically described in the Novogene NGS DNA Library Prep Kit, and sequencing was performed by Beijing Nuo Po-derived science and technology, inc.
(II) enrichment of X chromosome-specific DNA fragments
1. Obtaining the X chromosome specific candidate DNA fragment:
female fish reference genomes obtained before the laboratory (PRJNA 612372, femalees: SRR11300855, SRR11300854, SRR 11300853) and super male fish SM1-SM3 sequencing reads were aligned using bwa v0.7.17-r1188 (http:// bio-bw. Sourceforce. Net /) software, using a double-end alignment strategy, with base error rate set to 0.04, and the rest using default parameters. And (3) filtering contigs which can enable the super-male fish to sequence the reads with double ends aligned in the female fish reference genome, and selecting the contigs which cannot enable the super-male fish to sequence the reads with double ends aligned in the male fish reference genome, wherein the regions are possible X chromosome specific DNA fragments (figure 3).
2. Deep sequencing analysis of X chromosome specific fragments:
the candidate X-specific fragments obtained in the previous step were subjected to sequencing depth analysis and screening using Samtools (version 1.9). Reference genomes (PRJNA 612372, males: SRR11300858, SRR11300857, SRR11300856; females: SRR11300855, SRR11300854, SRR 11300853) of male (XY) and female (XX) mandarin fish previously obtained by the laboratory were selected for reference analysis, and in the X-chromosome and Y-chromosome sequencing depth comparison analysis, in order to avoid depth calculation abnormality caused by errors in sequence alignment, sequences with XY chromosome homology and similarity lower than 95% were selected for statistics in depth calculation. The screening principle is that the sequencing depth of the fragment is 0.3-0.7 times of the total depth of a male genome and 0.8-1.2 times of the total depth of a female genome, and the sequence conforming to the sequencing principle is a candidate X chromosome specific sequence. A total of 49X chromosome-specific Contig sequences meeting the above-mentioned screening conditions were obtained.
(III) screening of X chromosome molecular markers
1. Designing and synthesizing a primer:
6 of the Contig sequences were randomly selected based on the above 49X chromosome-specific Contig sequences, and PCR primers were designed using Primer5 software (the software used was the reference Primer PREMIER version5.0 for Windows and Power Macintosh) according to Contig-xxF/R (xx stands for Contig number), for example, contig-1 has Contig-1F/R (F stands for upstream Primer, R stands for downstream Primer, the same applies below). The primers were synthesized by Enwei fundi (Shanghai) trade Co., ltd.
2. Preparing a mandarin fish DNA sample:
shearing 4 female tail, 4 male tail and 4 super male individual tail fins of the mandarin fish during reproduction, numbering the female fish F1-F4, the male fish M1-M4 and the super male fish SM1-SM4 in sequence, preparing DNA samples by a conventional column type centrifugation method (a universal column type genome extraction kit, beijing kang is century biotechnology limited) according to the operation steps of a specification, and detecting the quality and the concentration by using 1% agarose gel electrophoresis and a micro spectrophotometer (NanoDrop 2000).
3. Screening and confirming of X chromosome molecular markers:
selecting the 6 pairs of primers to carry out PCR detection on 4 female fish, 4 male fish and 4 super male fish individuals, wherein the total PCR amplification reaction system is 20 mu L, including 10 mu L of Kangji 2 XTaq MasterMix (Beijing Kangji century Biotechnology Co., ltd.), each of the upstream and downstream primers (10 mu mol/L) is 0.8 mu L, the template DNA is 50ng, and the rest is ddH 2 And (4) supplementing and finishing.
The PCR reaction program is: pre-denaturation at 94 ℃ for 2min; denaturation at 94 ℃ for 30s, annealing at 56 ℃ for 30s, extension at 72 ℃ for 30s, and 35 cycles; final extension at 72 ℃ for 5min.
Agarose gel electrophoresis results show that 2 pairs of primers only obtain X chromosome specific amplification bands in male and female (XX, XY) individuals, no X chromosome specific amplification fragment is detected in super male (YY) individuals (figure 4), and the two pairs of primers are respectively Contig-1F/R and Contig-2F/R, which represents that the corresponding Contig is Contig-1 and Contig-2; the remaining 4 primer pairs, the same amplification products were present in female, male and supermale individuals (FIG. 5), demonstrating that these fragments are not X chromosome specific fragments.
2 of these contigs were confirmed by PCR amplification of 6 primer pairs in 4 individuals each of male, female and supermale: contig-1 and Contig-2 are X chromosome specific fragments, and are X chromosome molecular markers of mandarin fish.
Example 2
The embodiment provides a method for identifying a mandarin fish YY super-male fish by using a mandarin fish X chromosome molecular marker primer, which comprises the following steps:
(1) Preparing a siniperca chuatsi DNA sample:
visual inspection of the mandarin fish with mature nature is obtained from the Siniperca Chuatsi Miao GmbH of Fushan City of Guangdong province, 8 tails of each of female and male mandarin fish are obtained by dissection, and 6 tails of the super-male mandarin fish are obtained by test cross validation. Tail fins are respectively cut, and a conventional column type centrifugation method (a general column type genome extraction kit, beijing kang is century biotechnology limited) is also adopted to prepare DNA samples according to the operation steps of the instruction.
(2) And (3) PCR amplification verification:
PCR verification of the extracted mandarin fish DNA samples was performed using primers Contig-1F/R and Contig-2F/R (obtained in example 1).
The total PCR amplification reaction system is 20 μ L, including 10 μ L of 2 × Taq MasterMix (Beijing kang is century Biotechnology Co., ltd.), 0.8 μ L of each of the upstream and downstream primers (10 μmol/L), 50ng of template DNA, and ddH for the rest 2 And (4) supplementing by using oxygen.
The PCR reaction program is: firstly, pre-denaturation is carried out for 2min at 94 ℃; then carrying out denaturation at 94 ℃ for 30s, annealing at 56 ℃ for 30s and extension at 72 ℃ for 30s for 35 cycles; and final extension at 72 ℃ for 5min.
After the PCR reaction is finished, the product is subjected to agarose gel electrophoresis with the mass percentage of 1 percent, and the result is that: primers Contig-1F/R and Contig-2F/R can amplify bands in a DNA sample of the 16 normal male and female fish, and the molecular size of the X chromosome specific band amplified by the primers Contig-1F/R is about 935bp; the molecular size of the X chromosome-specific band amplified using Contig-2F/R was about 850bp. No X chromosome-specific band could be amplified in the 6-tailed super-male DNA samples (FIG. 6. The molecular markers Contig-1F/R and Contig-2F/R can be used for identifying the sex chromosome composition of male mandarin fish.
Example 3
The embodiment provides an application of the siniperca chuatsi X chromosome molecular marker primer in full male breeding of siniperca chuatsi, which comprises the following steps:
(1) After mandarin fish fries are cultured to 10 days old, dihydroestrone (E) with the concentration of 500mg/kg is added 2 ) Feeding bait fish to the eel and feeding the bait fish with hormone to the mandarin fish. Continuously feeding for two months, adopting natural illumination during the culture period, keeping the temperature of the culture water at 26-28 ℃, and feeding after being fed with satiety.
(2) XY pseudo female fish are screened by using Y chromosome molecular marker primers previously obtained in the laboratory (see application No. 201911100842.3), and then are mated with XY normal male fish, offspring fry are cultivated to 30 days old, offspring are screened by adopting the method in the example 2, and the super male fish with the genotype YY is obtained, and the specific process refers to the example 2.
(3) And (4) continuously culturing the YY super-male fish identified by the mark until the YY super-male fish is sexually mature, and mating the YY super-male fish with the normal XX female fish in a breeding season to obtain offspring which are the all-male fish.
The above embodiments are only used for illustrating the present invention, and the scope of the present invention is not limited to the above embodiments. The object of the present invention can be achieved by those skilled in the art based on the above disclosure, and any improvements and modifications based on the concept of the present invention fall within the protection scope of the present invention, which is described in the claims.
Claims (6)
1. A molecular marker primer of siniperca chuatsi X chromosome is characterized in that: the primer is a primer pair 1 or a primer pair 2, the primer pair 1 comprises a Contig-1 upstream primer and a Contig-1 downstream primer, and the primer pair 2 comprises a Contig-2 upstream primer and a Contig-2 downstream primer, wherein the nucleotide sequence of the Contig-1 upstream primer is shown in SEQ ID NO:1, the nucleotide sequence of the Contig-1 downstream primer is shown as SEQ ID NO:2, the nucleotide sequence of the Contig-2 upstream primer is shown as SEQ ID NO:3, the nucleotide sequence of the Contig-2 downstream primer is shown as SEQ ID NO:4, respectively.
2. A method for identifying YY super-male mandarin fish by tilting mouth is characterized by comprising the following steps:
(1) Preparing a siniperca chuatsi DNA sample;
(2) Carrying out PCR amplification by using the primer pair 1 or the primer pair 2 in the claim 1 by using the DNA sample in the step (1) as a template, carrying out electrophoresis detection after the amplification reaction is finished, wherein if the electrophoresis result shows that an amplification product has an X chromosome specific strip, the detected siniperca chuatsi DNA sample is XX female siniperca chuatsi or XY male siniperca chuatsi, and if the electrophoresis result shows that the amplification product does not have the X chromosome specific strip, the detected siniperca chuatsi DNA sample is YY super male siniperca chuatsi.
3. The method for identifying the YY super-male mandarin fish of claim 2, wherein the method comprises the following steps: in the step (2), the DNA sample in the step (1) is used as a template, a primer is adopted to carry out PCR amplification on the 1, electrophoresis detection is carried out after the amplification reaction is finished, if the electrophoresis result shows that an amplification product has an X chromosome specific strip of about 935bp, the detected siniperca chuatsi DNA sample is XX female siniperca chuatsi or XY male siniperca chuatsi, and if the electrophoresis result shows that the amplification product does not have the X chromosome specific strip of about 935bp, the detected siniperca chuatsi DNA sample is YY super-tip siniperca chuatsi.
4. The method for identifying the YY super-male mandarin fish of claim 2, wherein the method comprises the following steps: and (2) carrying out PCR amplification on the DNA sample in the step (1) serving as a template by using a primer pair 2, carrying out electrophoresis detection after the amplification reaction is finished, wherein if the electrophoresis result shows that an amplification product has an X chromosome specific strip of about 850bp, the detected DNA sample of the siniperca chuatsi is XX female siniperca chuatsi or XY male siniperca chuatsi, and if the electrophoresis result shows that the amplification product does not have the X chromosome specific strip of about 850bp, the detected DNA sample of the siniperca chuatsi is YY supermale chuatsi.
5. The use of the primer of claim 1 in whole male mandarin fish breeding.
6. The method of claim 2, wherein the method is applied to the whole male breeding of siniperca chuatsi.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116904474A (en) * | 2023-06-30 | 2023-10-20 | 中国长江三峡集团有限公司中华鲟研究所 | Male sex-specific molecular marker of siniperca scherzeri and genetic sex identification method |
| CN117660667A (en) * | 2024-02-02 | 2024-03-08 | 中山大学 | SNP molecular marker related to siniperca chuatsi growth trait and application thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116904474A (en) * | 2023-06-30 | 2023-10-20 | 中国长江三峡集团有限公司中华鲟研究所 | Male sex-specific molecular marker of siniperca scherzeri and genetic sex identification method |
| CN116904474B (en) * | 2023-06-30 | 2024-04-02 | 中国长江三峡集团有限公司中华鲟研究所 | Male sex-specific molecular marker of siniperca scherzeri and genetic sex identification method |
| CN117660667A (en) * | 2024-02-02 | 2024-03-08 | 中山大学 | SNP molecular marker related to siniperca chuatsi growth trait and application thereof |
| CN117660667B (en) * | 2024-02-02 | 2024-04-12 | 中山大学 | SNP molecular marker related to siniperca chuatsi growth trait and application thereof |
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