CN109161603A - The microsatellite marker primer and genetic sex identification method of channel catfish gunther sex-linked - Google Patents
The microsatellite marker primer and genetic sex identification method of channel catfish gunther sex-linked Download PDFInfo
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Abstract
The invention discloses the microsatellite marker primers and genetic sex identification method of channel catfish gunther sex-linked.Microsatellite marker primer and male unique allele sequence of the present invention according to the channel catfish gunther sex-linked of acquisition, create the PCR identification method of channel catfish genetic sex.The present invention screens the microsatellite marker and male unique allele of channel catfish gunther sex-linked for the first time, establishes the round pcr of channel catfish genetic sex identification.It is efficient, accurate and stable that the present invention has the characteristics that, has significant application value in channel catfish sexual control and unisexuality Seedling production, while also having important application prospect in clone's channel catfish sex determining gene and Sex Differentiation Mechanism Study.
Description
Technical field
The invention belongs to the molecular marking technique fields of genetics-breeding in fish, and in particular to a kind of to defend using fluorescent marker is micro-
Star primer identifies channel catfish heredity method for distinguishing.
Background technique
Channel catfish originates in the U.S., is a kind of economic species of worldwide aquatic products, introducing China in 1984.Because it has
Fine and tender taste, full of nutrition, dressing percentage is high, deep to be welcome by the majority of consumers the advantages that without piercing between flesh.Channel catfish dyeing
Body STUDY ON THE KARYOTYPE shows that diploid chromosome number mesh is 58,2n=58.It is marked and is constructed using different kinds of molecules such as AFLP, STR, SNP
Channel catfish linkage map, and combine chromosome band type analysis and whole-genome association as a result, showing that spot is pitched
Wei Channel-catfish is made of 28 pairs of autosomes and 1 pair of sex chromosome (respectively X and Y chromosome), belongs to XY/XX Sex Determination system,
Male is different to match, and female is same to match.Apparent dimorphism, i.e. the male speed of growth is presented in channel catfish male and female growth
With mature weight obviously higher than female individuals.Therefore, the molecule marking research of channel catfish gunther sex-linked is actively developed,
The molecular labeling with gunther sex-linked is excavated, channel catfish sexual control Mechanism Study, breeding population sex ratio will be controlled,
And all-male seed rearing etc. has important theory significance and application value.
Microsatellite marker is that have polymorphism height, stability strong, special by the short tandem repeat of 1-6 base composition
It is anisotropic high, and the characteristics of codominant inheritance is presented.Because it meets Mendelian inheritance law of segregation, homozygote can be accurately distinguished
And heterozygote.Genetic sex identification technology based on microsatellite parting is most widely used in current aquatic livestock pedigree confirmation
One of general most reliable means.In a variety of fish sex reversal research process, accurately identified using sex-kink microsatellite marker
Genetic sex is the female individuals of male out.Therefore, a set of quick, precise Identification spot based on microsatellite molecular marker is developed
Point fork tail Channel-catfish heredity method for distinguishing, the research of development and Sex Determination Mechanism to channel catfish breeding work have important
Meaning.
Summary of the invention
The object of the present invention is to provide a kind of allele that channel catfish male is special.
It is a further object of the present invention to provide a kind of microsatellite marker primers of channel catfish gunther sex-linked.
It is yet another object of the invention to provide a kind of channel catfish microsatellite genetic sex identification methods, are channel catfish
The research of Channel-catfish Sex Determination Mechanism, all-male seed rearing provide basis.Mainly pass through channel catfish gunther sex-linked microsatellite mark
Remember that site carries out PCR amplification and the detection of Capillary Electrophoresis parting to realize.
The technical solution that the invention patent is taken:
A kind of allele that channel catfish male is special, as shown in SEQ ID NO.27.
Application of the allele of the present invention as detection target spot in channel catfish genetic sex identification.
The application expands channel catfish preferably by special primer shown in SEQ ID NO.1 and SEQ ID NO.2
Genomic DNA, the individual that can amplify the special allele of the male of channel catfish shown in SEQ ID NO.27 is male
Individual.
A kind of microsatellite marker primer of channel catfish gunther sex-linked, by 2 special primer SEQ ID NO.1 and SEQ
ID NO.2 composition, wherein 5 ' the ends of SEQ ID NO.1 are marked through FAM fluorophor.
Application of the microsatellite marker primer of the present invention in channel catfish genetic sex identification.
A kind of method of channel catfish genetic sex identification, extracts the channel catfish genome of genetic sex to be measured
DNA, with the primer SEQ ID NO.1 and normal primer SEQ ID NO.2 marked through FAM fluorophor to channel catfish gene
Group DNA carries out PCR amplification, and amplified production carries out Capillary Electrophoresis, LIZ-500 conduct on ABI 3730XL genetic analysis systems
Internal reference reads amplified production genotype with GeneMarker v2.2.0 software；It is with size on Capillary Electrophoresis map
The individual of the DNA fragmentation of 186bp is the channel catfish that genetic sex is male, and the heredity of the individual without this allele C
Gender is female.
The utility model has the advantages that
The microsatellite marker of channel catfish gunther sex-linked has been screened for the first time, it was found that the special allele of male,
Establish the genetic sex identification method based on micro-satellite labeling technique.It is efficient, accurate and stable that the present invention has the characteristics that,
Channel catfish sexual control, seed rearing early sex identification and all-male Seedling production in have significant application value.
Detailed description of the invention
Fig. 1 iso-allele base sequence for the positioning of channel catfish microsatellite locus JSCC09128 genome and not.
Fig. 2 is channel catfish microsatellite locus JSCC09128 genotypic results.A:AC type, B:BC type, A and B are hero
Property genes of individuals genotyping result；C:AA type, D:AB type, E:BB type, C, D and E are female individuals genotypic results.
Specific embodiment
Embodiment 1
It includes three sides that the present invention, which is described in detail, to the technology of the present invention content below by way of examples and with reference to the accompanying drawings
Face: (1) screening of channel catfish gunther sex-linked microsatellite marker and its primer sequence；(2) channel catfish gunther sex-linked
The verifying of microsatellite marker；(3) application method of the channel catfish genetic sex identification based on microsatellite marker.
1. screening and its primer sequence of channel catfish gunther sex-linked microsatellite marker
1.1. the screening of gunther sex-linked microsatellite
156 tail of channel catfish family full-sibs individual and Parent DNA sample are acquired, constructs 19 libraries, sequencing altogether
Data are 147Gb, and Parent sequencing data is 3Gb, covering gene group is about 3 ×, the average data of each sample is sequenced in filial generation
Amount be 900Mb, covering gene group be 1 ×.Using preliminary screening to 10661 SNP carry out point group and genetic linkage maps structure
It builds, LOD > 10, separates 29 linkage groups.Reference numerals eventually for building genetic linkage maps are 5017, and genetic distance is long
Degree is 3147.3cM, average length 0.63cM.High quality microsatellite marker progressive linkage analysis is screened, 13 is obtained altogether and dives
The 17th linkage group of channel catfish is navigated in sex-kink microsatellite marker.
1.2. the design of micro-satellite primers
13 microsatellite markers of acquisition are accurately positioned and arrive its genome position.It is set with the online primer of primer 3
The flanking sequence design primer of software needle microsatellite marker is counted, the raw work biology in primer synthesis commission Shanghai carries out.
1 channel catfish gunther sex-linked microsatellite locus of table and primer information
Note: F is that forward primer R is reverse primer
2. the verifying of the microsatellite marker of gunther sex-linked
2.1. extracting genome DNA
The channel catfish male and each 16 tail of female individuals of known gender, the every tail channel catfish tail of clip are chosen respectively
Fin sample about 25mg is transferred in 1.5ml centrifuge tube with liquid nitrogen grinding at powder.It is slow that 450 μ L STE DNA extracting is successively added
Fliud flushing (10mmol/L Tris-HCl, pH 8.0；1mmol/L EDTA, pH 8.0), 35 μ L SDS (10%), be eventually adding 15 μ
L Proteinase K (0.2%) is mixed by inversion.RnaseA1 μ L is added, is placed in 55 DEG C of warm bath 1h of water-bath.Isometric (about 700 μ are added
L) Tris saturated phenol is vibrated on DNA mixed instrument and is mixed, and 12000rpm is centrifuged 10min at 4 DEG C, after will using liquid-transfering gun
Supernatant is transferred in another clean centrifuge tube.Isometric phenol is added in supernatant, and to imitate alcohol mixture (phenol, chloroform, different
Amylalcohol ratio is 25:24:1), it is placed in oscillation on DNA mixed instrument and mixes 15min.Isometric chloroform about 500 is added in supernatant
μ L is placed in oscillation on DNA mixed instrument and mixes 15min.The dehydrated alcohol 1mL that -20 DEG C of pre-coolings are added in supernatant precipitates DNA,
Supernatant is abandoned after centrifuge 12000rpm centrifugation 5min.70% ethanol washing is first added twice, adds dehydrated alcohol washing one
It is secondary, 200 μ L TE are added after dry, dissolution is abundant.With NanoDrop ND-1000 spectrophotometer detectable concentration, and by each DNA
Sample is diluted to the working solution of 100ng/ μ L.
2.2.PCR amplification
Each pair of micro-satellite primers carry out PCR amplification to 16 tail males and female sample respectively.PCR amplification uses 2 × Taq
Plus Master Mix (Dye Plus) kit (Nanjing Vazyme Biotechnology Co., Ltd.), reaction system are as follows:
Amplification condition are as follows: 94 DEG C of initial denaturation 5min；94 DEG C of denaturation 30s, 60 DEG C~51 DEG C (each circulation subtract 1 DEG C) annealing
30s, 72 DEG C of extension 60s, each annealing temperature 2 circulations；94 DEG C of denaturation 30s, 55 DEG C of annealing 30s, 72 DEG C of extension 60s, 15
Circulation；72 DEG C of extension 10min.
2.3. polyacrylamide gel electrophoresis
Above-mentioned pcr amplification product carries out electrophoresis, silver staining on 6% denaturing polyacrylamide gel.Silver staining step is such as
Under: it 1. fixes, the glass plate for being stained with gel is immersed in acetic acid aqueous solution about 10min；2. rinsing, glass plate is transferred to distilled water
In rinse acetic acid；3. dyeing, 30min is impregnated in dyeing liquor (0.2%AgNO3 100mL adds 37% formaldehyde, 50 μ L)；4. again
Secondary rinsing rinses about 5s in distilled water；5. developing the color, rinsed gel glass plate is transferred to developing solution (1.5%NaOH immediately
100mL adds 37% formaldehyde 0.5mL), and shake on shaking table until aobvious band；6. glass plate is transferred to acetic acid and fixed by color development stopping
In liquid, color development stopping reaction.Then it dries and saves and taken a picture with digital camera.Compare microsatellite allele in male and female individual
In distribution whether there were significant differences.Finally screen the microsatellite mark that primer sequence is SEQ ID NO.1 and SEQ ID NO.2
Remember JSCC09128 and channel catfish gender close linkage.
2.4. differential fragment is cloned
It is female to further use sexually matured 20 tail of channel catfish of primer SEQ ID NO.1 and SEQ ID NO.2PCR amplification
Fish and 20 tail milter genomic DNAs, PCR amplification use the kit (Nanjing 2 × Taq Plus Master Mix (Dye Plus)
Nuo Weizan Biotechnology Co., Ltd), reaction system are as follows:
Amplification condition are as follows: 94 DEG C of initial denaturation 5min；94 DEG C of denaturation 30s, 60 DEG C~51 DEG C (each circulation subtract 1 DEG C) annealing
30s, 72 DEG C of extension 60s, each annealing temperature 2 circulations；94 DEG C of denaturation 30s, 55 DEG C of annealing 30s, 72 DEG C of extension 60s, 15
Circulation；72 DEG C of extension 10min.
Amplified production gel extraction simultaneously connects 2h under the conditions of 16 DEG C with PMD-18T carrier (Takara) after purification.It will be even
Object of practicing midwifery, which is added to, to be cloned and is coated in competent cell on the LB plate containing acillin, and 37 DEG C of inversions were cultivated
Night.10 positive colonies are selected, in the fluid nutrient medium containing ampicillin, 37 DEG C of shake culture 5h are taken out using plasmid
Extraction reagent kit carries out Sanger sequencing after extracting plasmid.Sanger sequencing result is read using Chromas software.
SEQ ID NO.1 and SEQID NO.2 primer amplifies 3 allele, sequence length in 20 male samples
Respectively 192bp, 189bp, 186bp are respectively designated as allele A, B, C according to size order, by this section of sequence in spot
It is positioned on fork tail Channel-catfish genome, the results showed that the microsatellite marker is located at second introne of ZBTB38-like gene
Region, genome positioning and allele partial sequence are as shown in Figure 1.Allele is only amplified in 20 female samples
A and B.Sequencing result shows that, relative to allele A, there are the missings of 3 bases (to lack 1 in the duplicate block TAA by allele B
A TAA is repeated), there are the missing of 6 bases (lacking 2 TAA to repeat) in the duplicate block TAA for allele C.Therefore C equipotential base
Because that may be the special allele of channel catfish male.Supposition allele C is the distinctive allele of male.
2.4. channel catfish male unique allele is verified
96 tails individual is selected at random in channel catfish breeding population, with what is marked through fluorophor FAM (blue)
SEQID NO.1 sequence and the combination of SEQID NO.2 aligning primer carry out PCR amplification.Amplified production is in ABI 3730XL heredity point
Capillary Electrophoresis is carried out on analysis system, uses LIZ-500 as internal reference, reads each sample with GeneMarker v2.2.0 software
Genotype.
Using 2 × Taq Plus Master Mix (Dye Plus) kit, (biotechnology is only praised in Nanjing promise to PCR amplification
Co., Ltd), reaction system are as follows:
Amplification condition are as follows: 94 DEG C of initial denaturation 5min；94 DEG C of denaturation 30s, 60 DEG C~51 DEG C (each circulation subtract 1 DEG C) annealing
30s, 72 DEG C of extension 60s, each annealing temperature 2 circulations；94 DEG C of denaturation 30s, 55 DEG C of annealing 30s, 72 DEG C of extension 60s, 15
Circulation；72 DEG C of extension 10min.
Genotyping result is compared with counter sample record gender in database, shows expand in all males
Increase the DNA fragmentation that size out is 186bp, and without this amplified production in female individuals, illustrate to turn by male unique allele
The molecular labeling of change can accurately identify the genetic sex of channel catfish.
The application method of channel catfish genetic sex identification of the embodiment 2 based on microsatellite marker
1. extracting genome DNA
From channel catfish G12011 grades of breeding populations of generation and 2013 grades of breeding populations select 32 tails and 64 tail parents at random
Sample, extracting genome DNA is the same as 2.1.
2. fluorescent primer synthesizes
SEQ ID NO.1 primer sequence is synthesized, in its 5 ' end addition FAM fluorescence light group (blue), SEQID NO.2 sequence
Column do not make fluorescence addition processing, and primer synthesis commission Shanghai Sheng Gong biotech firm completes.
3.PCR amplification
Using the unknown channel catfish genomic DNA of above-mentioned 96 tail gender as template, with the SEQ of addition FAM fluorophor
ID NO.1 and SEQID NO.2 primer carries out PCR amplification, and PCR amplification uses 2 × Taq Plus Master Mix (Dye
Plus) kit (Nanjing Vazyme Biotechnology Co., Ltd.), reaction system are as follows:
Amplification condition are as follows: 94 DEG C of initial denaturation 5min；94 DEG C of denaturation 30s, 60 DEG C~51 DEG C (each circulation subtract 1 DEG C) annealing
30s, 72 DEG C of extension 60s, each annealing temperature 2 circulations；94 DEG C of denaturation 30s, 55 DEG C of annealing 30s, 72 DEG C of extension 60s, 15
Circulation；72 DEG C of extension 10min.
4. Genotyping
Amplified production carries out Capillary Electrophoresis on ABI 3730XL genetic analysis systems, uses GS-500LIZ as internal reference.
Capillary electrophoresis test system is 20 μ L: wherein Hi-Di Formamide and 10 μ L of LIZ-500 mixture, pcr amplification product 10
μL。
Deposition condition are as follows: voltage 15KV, capillary pipe length 50cm, 60 DEG C of electrophoresis temperature, time 2.5h.
Amplified production genotype is read with GeneMarker v2.2.0 software.
5. interpretation of result
After determining respectively 96 tail channel catfish microsatellite locus JSCC09128 genotype, genetic sex supposition is carried out.
The result shows that: genetic sex is female sample totally 53 tail, and genotype is 33 tail of sample of AA, 14 tail of AB, 6 tail of BB；Heredity
43 tail of sample of gender tail male, genotype are 38 tail of sample of AC, 5 tail of BC.The corresponding capillary electricity of every kind of genotype combination
Map of swimming is as shown in Figure 2.Counter sample in the channel catfish sample gender speculated according to Genotyping and database is recorded
Gender checked, as a result 100% matching.
Sequence table
<110>Jiangsu Fresh Water Aquatic Product Inst.
<120>the microsatellite marker primer and genetic sex identification method of channel catfish gunther sex-linked
<160> 27
<170> SIPOSequenceListing 1.0
<210> 1
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 1
tgaatgtgag actaacagga g 21
<210> 2
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
acatcgcttt gagaagctgc t 21
<210> 3
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 3
ggccgcacaa ttcgaatagt 20
<210> 4
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 4
agccagtctc tctacatgca 20
<210> 5
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 5
tctcatactg gctgtgctgt 20
<210> 6
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 6
cggaatcgca tagagggtta a 21
<210> 7
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 7
accagcttta cctcgtttac c 21
<210> 8
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 8
agtgttctga tttgtgtggc t 21
<210> 9
<211> 23
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 9
gtcttggagt ctatttggaa cca 23
<210> 10
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 10
cagacggaca catggacaac 20
<210> 11
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 11
ttagccttga cctgtggaca 20
<210> 12
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 12
cgatgcatcc atgtacaggc 20
<210> 13
<211> 22
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 13
agggtatttg tctctccttg ct 22
<210> 14
<211> 19
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 14
acacaccgat cagactcaa 19
<210> 15
<211> 22
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 15
tcctgtagtc attggtgtgt ga 22
<210> 16
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 16
gcctgttagc atcttcagcc 20
<210> 17
<211> 22
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 17
agaaatggtg ctgtgtgatg ag 22
<210> 18
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 18
accccatcct gtgttttgtt 20
<210> 19
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 19
cccttaaccc tctctgctcc 20
<210> 20
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 20
tgtagaaccc acccttttcc t 21
<210> 21
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 21
gttcagcttt atcggccagg 20
<210> 22
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 22
cagaggtgtt tggtggctac 20
<210> 23
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 23
tgctgtgtct gaattccgga 20
<210> 24
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 24
tcaactttgt ggcagcagtt 20
<210> 25
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 25
tcgatcgaaa gtgggcaaac 20
<210> 26
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 26
cactccaacg ggctctctag 20
<210> 27
<211> 186
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 27
tgaatgtgag actaacagga ggtttttttt tagaattcag tcaatgtgaa tatgaattaa 60
taacattcaa taagttaaga aatcttactt taatcttcag aatttagtta ataataataa 120
ttaataataa taataataag ctttatttta taaagcgcct ttaaaagcag cttctcaaag 180
cgatgt 186
Claims (6)
1. a kind of allele that channel catfish male is special, it is characterised in that as shown in SEQ ID NO.27.
2. application of the allele described in claim 1 as detection target spot in channel catfish genetic sex identification.
3. application according to claim 2, it is characterised in that utilize special shown in SEQ ID NO.1 and SEQ ID NO.2
Primer amplification channel catfish genomic DNA, can amplify the male of channel catfish shown in SEQ ID NO.27 it is special etc.
The individual of position gene is male.
4. a kind of microsatellite marker primer of channel catfish gunther sex-linked, it is characterised in that by 2 special primer SEQ ID
NO.1 and SEQ ID NO.2 composition, wherein 5 ' the ends of SEQ ID NO.1 are marked through FAM fluorophor.
5. application of the microsatellite marker primer as claimed in claim 4 in channel catfish genetic sex identification.
6. a kind of method of channel catfish genetic sex identification, it is characterised in that extract the channel catfish of genetic sex to be measured
Genomic DNA, with the primer SEQ ID NO.1 and normal primer SEQ ID NO.2 marked through FAM fluorophor to channel catfish
Channel-catfish genomic DNA carries out PCR amplification, and amplified production carries out Capillary Electrophoresis, LIZ- on ABI 3730XL genetic analysis systems
500 are used as internal reference, read amplified production genotype with GeneMarker v2.2.0 software；Have on Capillary Electrophoresis map big
It is small be the individual of the DNA fragmentation of 186bp be genetic sex be male channel catfish, and without this allele C individual
Genetic sex is female.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111304338A (en) * | 2020-03-13 | 2020-06-19 | 江苏省淡水水产研究所 | SNP molecular marker linked with sex of channel catfish and genetic sex identification method |
| CN112342214A (en) * | 2020-11-11 | 2021-02-09 | 江苏省淡水水产研究所 | sgRNA sequence for targeted knockout of channel catfish zbtb38 gene and screening method thereof |
| CN113186307A (en) * | 2021-06-21 | 2021-07-30 | 江苏省淡水水产研究所 | Sex-linked SNPs (single nucleotide polymorphisms) marker development method based on channel catfish male specific gene zbtb38-Y and application |
| CN113293218A (en) * | 2021-06-21 | 2021-08-24 | 江苏省淡水水产研究所 | SNP molecular marker for selecting weight gain character of channel catfish and application |
| CN113373244A (en) * | 2021-07-14 | 2021-09-10 | 江苏省淡水水产研究所 | Method for rapidly detecting genetic sex of channel catfish based on SNP site-specific primer extension reaction |
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| CN111304338A (en) * | 2020-03-13 | 2020-06-19 | 江苏省淡水水产研究所 | SNP molecular marker linked with sex of channel catfish and genetic sex identification method |
| CN111304338B (en) * | 2020-03-13 | 2023-03-31 | 江苏省淡水水产研究所 | SNP molecular marker linked with sex of channel catfish and genetic sex identification method |
| CN112342214A (en) * | 2020-11-11 | 2021-02-09 | 江苏省淡水水产研究所 | sgRNA sequence for targeted knockout of channel catfish zbtb38 gene and screening method thereof |
| CN112342214B (en) * | 2020-11-11 | 2024-03-26 | 江苏省淡水水产研究所 | sgRNA sequence of targeted knockout channel catfish zbtb38 gene and screening method thereof |
| CN113186307A (en) * | 2021-06-21 | 2021-07-30 | 江苏省淡水水产研究所 | Sex-linked SNPs (single nucleotide polymorphisms) marker development method based on channel catfish male specific gene zbtb38-Y and application |
| CN113293218A (en) * | 2021-06-21 | 2021-08-24 | 江苏省淡水水产研究所 | SNP molecular marker for selecting weight gain character of channel catfish and application |
| CN113186307B (en) * | 2021-06-21 | 2023-08-01 | 江苏省淡水水产研究所 | Sex-linked SNPs (single nucleotide polymorphisms) marker development method and application based on channel catfish male specific gene zbtb38-Y |
| CN113293218B (en) * | 2021-06-21 | 2024-02-06 | 江苏省淡水水产研究所 | SNP molecular marker for selecting weight gain character of channel catfish and application |
| CN113373244A (en) * | 2021-07-14 | 2021-09-10 | 江苏省淡水水产研究所 | Method for rapidly detecting genetic sex of channel catfish based on SNP site-specific primer extension reaction |
| CN113373244B (en) * | 2021-07-14 | 2023-05-23 | 江苏省淡水水产研究所 | Method for rapidly detecting genetic sex of channel catfish based on SNP locus specific primer extension reaction |
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