CN112980962A - SNP marker related to birth weight trait of pig and application thereof - Google Patents
SNP marker related to birth weight trait of pig and application thereof Download PDFInfo
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Abstract
The application discloses an SNP marker related to the birth weight of pigs and application thereof. The SNP marker related to the birth weight trait of the pig is located at the 112031589 th nucleotide site on the No. 4 chromosome of a reference sequence of the 11.1 edition of the pig international genome, and has G or A polymorphism. The SNP marker related to the birth weight character of the pig can be used for the birth weight prediction of piglets and the molecular marker-assisted selection and breeding of the pig. By adopting the SNP marker for breeding, the breeding time can be shortened, and the breeding accuracy can be improved; and is not easily influenced by other external environments and other factors; the piglet early weight can be effectively improved, so that the survival rate, the weaning weight and the fattening effect are improved, and the piglet early weight feed has great significance for the breeding production of pigs. By adopting the SNP marker of the application to assist breeding, early breeding can be carried out, timely elimination can be realized, the breeding cost can be reduced, homozygous parents can be quickly obtained, and the breeding efficiency and quality can be improved.
Description
Technical Field
The application relates to the technical field of pig breeding, in particular to an SNP marker related to the birth weight trait of a pig and application thereof.
Background
Pork is rich in protein, vitamins and minerals and is a main non-staple food in daily life of residents in China. Although the population growth speed of China is slowing down, the population number is still in a continuously increasing state, and the demand for pork is kept in a growing state. Therefore, the breeding industry needs to improve the supply capacity of high-quality pork.
The weight of a piglet at birth, namely the birth weight of the piglet, is an important quantitative character for measuring the piglet and influences the growth and development, the production performance, the carcass grade and the like of the piglet in the whole life. The birth weight of the pig is positively correlated with the slaughter weight of the pork, and is negatively correlated with the slaughter age in days. According to the report, most piglets with the birth weight of less than 1.0kg have poor growth and development, and the piglets with the birth weight of more than 1.5kg have obviously better development. Researches show that positive correlations exist between the birth weight of piglets and the survival rate of weaning, the weight of weaning and the weight of slaughtering; when the birth weight of the piglet is increased by 0.1kg, the weaning weight is increased by 1kg, and the slaughtering weight is increased by 10 kg. Therefore, the method can improve the birth weight of the piglets in a targeted manner, and the piglets can have higher survival rate and better production performance, thereby bringing greater economic benefit to producers.
The birth weight of piglets is a complex quantitative character, is regulated and controlled by multiple genes together, and is influenced by various factors such as variety, birth times, nutrition, environment and the like. The traditional breeding method mostly adopts the traditional phenotype breeding technology, namely, the productivity of the parents is deduced according to the phenotype information of the offspring, the breeding period is long, and the genetic progress is slow. The molecular Marker Assisted Selection (MAS) is adopted for breeding, and the method has the advantages that the influence of other external environments and other factors is not easy to influence, the breeding time can be shortened, and the breeding accuracy is improved. The MAS technology is utilized to improve the birth weight character of the boar, the bigger the birth weight of the piglet is, the higher the survival rate is, the bigger the weaning weight is, the better the fattening effect is, and the MAS technology has great significance for improving the economic benefit in the breeding production.
Therefore, intensive research and excavation on genes and molecular markers influencing the birth weight of pigs are needed to meet the requirements of molecular marker-assisted selective breeding of the pigs.
Disclosure of Invention
The application aims to provide a novel SNP marker related to the birth weight trait of pigs and application thereof.
The application specifically adopts the following technical scheme:
the first aspect of the application discloses a SNP marker related to the birth weight trait of a pig, wherein the SNP marker is positioned at the 112031589 th nucleotide site on the No. 4 chromosome of a reference sequence of the 11.1 edition of the pig international genome and has a G or A polymorphism.
It should be noted that, by studying 139634 SNP markers of 674 sows, the 112031589 nucleotide site on chromosome 4 of the sow is finally related to the pig birth weight of piglets born by the sow. The research shows that the polymorphism of the SNP site is G or A, and the birth weight of piglets born by sows with AA genotype is obviously larger than that of sows with AG genotype or GG genotype.
Preferably, the SNP marker of the present application is located at base 127 from the 5' end of the sequence shown in SEQ ID No. 1.
Note that the 127 th base of the sequence shown in SEQ ID No.1 is only for the purpose of displaying the SNP marker of the present application in a simpler and clearer manner. The sequence shown in SEQ ID No.1 is only an amplification sequence of the SNP marker detection primer specifically designed in one implementation manner of the application; it will be appreciated that if a different detection primer is used, the SNP marker of the present application may not be located at base 127; however, if the reference sequence is found in version 11.1 of the porcine international genome, the exact location of the SNP marker of the present application is at nucleotide 112031589 on chromosome 4.
Preferably, the SNP marker of the present application is a sow having a piglet with a significantly greater birth weight than a sow with an AG genotype or a GG genotype of a sow with an AA genotype.
The second aspect of the application discloses a primer pair for detecting the SNP marker of the application, wherein the upstream primer of the primer pair is a sequence shown in Seq ID No.2, and the downstream primer is a sequence shown in Seq ID No. 3;
Seq ID No.2：5’-TCTTCGGGTCAGAAAGAGA-3’
Seq ID No.3：5’-CTTCCTGCAGATATTGGGG-3’。
the primer set having the sequence shown in Seq ID No.2 and the sequence shown in Seq ID No.3 is only one embodiment of the present application, and the SNP marker of the present application can be included therein; it can be understood that, on the one hand, on the basis of the primer pair of the present application, one skilled in the art can increase or decrease several bases at the 5 'end or 3' end thereof as required, as long as PCR amplification is not affected; on the other hand, the SNP marker of the present application does not exclude the possibility of designing another primer set as long as the SNP marker of the present application can be included in the amplified target sequence and the SNP marker detection of the present application can be achieved.
In a third aspect of the present application, a kit for detecting the SNP marker of the present application is disclosed, which comprises the primer pair of the present application.
Preferably, the kit of the present application further comprises reagents for PCR amplification.
The primer set of the present application can be used for detecting the SNP marker of the present application, and therefore, it can be assembled into a kit for detecting the SNP marker of the present application for the convenience of use; in principle, all reagents required for the detection, such as PCR amplification reagents, can be included in the kit for ease of use; however, it is understood that reagents other than the primer pairs of the present application can be purchased directly; therefore, the kit of the present application may be assembled with other reagents according to the product design requirements, except for the primer pair of the present application, and is not specifically limited herein.
The fourth aspect of the application discloses a method for detecting the SNP marker of the application, which comprises the steps of carrying out PCR amplification on the genomic DNA of a sow to be detected by adopting the primer pair of the application or the kit of the application, and obtaining the base condition of the SNP marker of the application by determining the sequence of a PCR amplification product.
It is understood that the PCR amplification of the genomic DNA and the sequencing of the amplified product to obtain the base information of the SNP marker is a more conventional and more accurate way to obtain the base information of the SNP marker, and does not exclude that the base information of the SNP marker of the present application can also be obtained by other ways, for example, after the PCR amplification, the mass spectrometry analysis of the amplified product to obtain the base information of the SNP marker of the present application, which is not limited herein.
The fifth aspect of the application discloses a method for predicting the pig birth weight of a piglet, which comprises the steps of carrying out PCR amplification on the genomic DNA of a sow by adopting the primer pair or the kit, obtaining the base condition of the SNP marker of the application by determining the sequence of a PCR amplification product, and predicting the pig birth weight of the piglet produced by the sow according to the base condition of the SNP marker of the application.
It should be noted that, the research in the present application shows that the birth weight of piglets born by sows with AA genotype is significantly larger than that of sows with AG genotype or GG genotype; therefore, by adopting the method for predicting the pig birth weight of the piglet, the pig birth weight of the piglet can be predicted approximately by detecting the genotype of the SNP marker of the sow. However, it can be understood that the pig birth weight of piglets is a complex quantitative trait, and is influenced by various factors such as breed, birth times, nutrition, environment and the like besides the genotype of the SNP marker of the application; therefore, the method for predicting the pig birth weight of the piglet, on one hand, only predicts and evaluates the pig birth weight of the piglet of the sow under the same or similar conditions; on the other hand, the method for predicting the pig birth weight of the piglets is only statistically significant, has important prediction on pig breeding, and does not accurately predict the specific weight of the pig birth weight of each piglet.
The sixth aspect of the application discloses the application of the SNP marker in pig birth weight prediction or pig molecular assisted screening breeding.
It is understood that the studies of the present application show that the SNP markers of the present application are closely related to the birth weight trait of swine, and thus, can be used for the prediction of the birth weight of swine or the molecular-assisted screening breeding of swine. Compared with the traditional breeding method, the molecular assisted screening breeding of the pigs is carried out by adopting the SNP marker, so that the breeding time can be shortened, the breeding accuracy is improved, and the influence of other factors such as external environment and the like is not easy to occur; by adopting the SNP marker, the MAS technology is utilized to improve the birth weight character of the boar and improve the birth weight of the piglet, thereby improving the survival rate, the weaning weight, the fattening effect and the like, and having great significance for improving the economic benefit of breeding production.
The sixth aspect of the application discloses the application of the primer pair or the kit in pig birth weight prediction or pig molecular assisted screening and breeding.
It can be understood that the primer pair and the kit of the present application are used for detecting the SNP marker of the present application, and the SNP marker of the present application is closely related to the birth weight trait of the pig, so the primer pair and the kit of the present application can be used for pig birth weight prediction or pig molecular assisted screening breeding.
The beneficial effect of this application lies in:
the SNP marker related to the birth weight character of the pig can be used for the birth weight prediction of piglets and the molecular marker-assisted selection and breeding of the pig. By adopting the SNP marker for breeding, the breeding time can be shortened, and the breeding accuracy can be improved; and is not easily influenced by other external environments and other factors; the piglet early weight can be effectively improved, so that the survival rate, the weaning weight and the fattening effect are improved, and the piglet early weight feed has great significance for the breeding production of pigs. By adopting the SNP marker of the application to assist breeding, early breeding can be carried out, timely elimination can be realized, the breeding cost can be reduced, homozygous parents can be quickly obtained, and the breeding efficiency and quality can be improved.
Drawings
FIG. 1 is a Manhattan plot of SNP markers significantly associated with swine birth weight status in the examples of the present application;
FIG. 2 is a partial agarose gel electrophoresis of the amplification products of PCR amplification of the genomic DNA of sows of three genotypes using primers designed in the examples of this application;
FIG. 3 is a partial sequencing result of the amplification product obtained by PCR amplification of the genomic DNA of a sow of three genotypes using primers designed in the examples of the present application.
Detailed Description
Compared with the traditional breeding mode of deducing parent productivity according to offspring phenotype information, the MAS technology can carry out typing identification on candidate groups according to positioned significant relevant sites, can carry out early breeding and timely eliminate, reduces breeding cost, quickly obtains homozygous parents and improves breeding progress. However, not all traits have significant associated loci, or not all traits can effectively correspond to or develop associated SNP loci for use in typing identification and molecular marker assisted breeding.
In the process of carrying out a large amount of research and experiments on the birth weight of piglets, the 112031589 th nucleotide site on the No. 4 chromosome of a reference sequence of the version 11.1 of the international genome of the pigs is found to be closely related to the birth weight of the piglets, and the research shows that the site has G or A polymorphism, and the birth weight of the piglets produced by the sows with AA genotype is obviously larger than that of the sows with AG genotype or GG genotype. Based on the above research, the present application proposes a SNP marker associated with the birth weight trait of swine, i.e., 112031589 th nucleotide site on chromosome 4, i.e., 127 th nucleotide from 5' end of the sequence shown in SEQ ID No.1 of the present application.
The SNP marker is closely related to the birth weight of the pig, and the detection of the SNP marker can predict the birth weight of the piglet, and can also be used for molecular marker assisted breeding of the pig and breeding of AA genotype sows, so that the birth weight of the piglet is improved, the survival rate of the piglet is improved, the weaning weight is increased and the fattening effect is improved. By adopting the SNP marker of the application to breeding, early breeding can be carried out, timely elimination can be realized, the breeding cost is reduced, homozygous parents can be quickly obtained, and the breeding efficiency and quality are improved.
The present application will be described in further detail with reference to specific examples. The following examples are intended to be illustrative of the present application only and should not be construed as limiting the present application.
Examples
Acquisition of porcine birth weight related SNP (single nucleotide polymorphism) sites
In the example, 674 sows are selected in total, wherein 453 Dabai pigs and 221 Changbai pigs are selected, and the weight of the offspring piglets is weighed for 2 hours through a standard weight scale to be used as the birth weight shape and form value of the pigs. The study of the SNP sites related to the birth weight of the pig is as follows:
1. genomic DNA extraction
A tissue sample with the size of soybeans is collected and stored in 75% ethanol for later use. The DNA extraction adopts a blood/tissue DNA magnetic bead extraction kit with the product number of GO-BTCD-400, and is carried out according to the standard flow of the instruction. The extracted DNA was subjected to restriction of Invitrogen corporationTMThe dsDNAHS Assay Kit was used for quantification and stored at-20 ℃.
2. Pig whole genome sequencing typing
By utilizing a Restriction-site associated DNA sequencing technology (RAD-seq), a library is built according to the standard flow of the MGIEasy simplified genome library preparation kit, and double-end sequencing (PE100) is carried out on the genome DNA extracted from the pig ear tissue sample by adopting a BGISEQ-500 sequencing platform.
After the sequencing data is downloaded, extracting individual data according to the tag sequence, filtering out low-quality sequencing fragments, and then comparing the high-quality fragments to an international pig reference genome 11.1 version (Sscrofa 11.1) by adopting BWA software (Li, Durbin, 2009); then, mutation detection was performed using gatk, which initially yielded 4199546 SNP markers from the genomic DNA sequencing results of 674 sows.
For the preliminary obtained 4199546 SNP markers, according to the hard filtering conditions: 'QUAL <30| | QD <2.0| | FS >60.0| | MQ <40.0| | MQRankSum < -12.5| | ReadPosRankSum < -8.0' performs hard filtration to remove false positive sites, and 3949591 SNP markers with higher quality are obtained.
Vcftools software was used, according to a minimum allele frequency of 0.05(maf 0.05), a SNP detection rate of 0.5(SNP call rate 0.5), and a hardy weinberg equilibrium coefficient of 10-6(hwe＝10-6) Quality control standard ofAnd (3) carrying out quality control, and finally, further screening to obtain 140948 SNP markers.
Filling the deletion genotype by using Beagle software, and performing secondary quality control according to the same quality control standard after filling to finally obtain 139634 high-quality SNP markers for later use.
3. And (3) carrying out whole gene association analysis on the birth weight of the pig:
GWAS analysis was performed on sow birth weight using EMMAX program (http:// genetics. cs. ucla. edu/EMMAX/index. html). The analytical model is as follows:
y＝Xb+Zu+Mg+e
in the model, y represents a real value of the character record; x represents a fixed effect correlation matrix; b represents a fixed effect vector; fixed effects include breed, year-season and birth; z represents an additive genetic effect correlation matrix; u represents an individual additive genetic effect vector, obeying a normal distribution,
m represents an SNP marker effect incidence matrix; g represents SNP marker effect; e denotes the residual, following a normal distribution,
g represents a genome genetic relationship matrix; i represents an identity matrix;
and
additive genetic effect variance and residual variance are indicated, respectively.
GWAS results show that the SNP marker related to the porcine birth weight trait is located at the 112031589 th nucleotide site on the No. 4 chromosome of the reference sequence of the 11.1 version of the porcine international genome, has a G/A polymorphism (g.112031589G/A), and reaches the chromosome significance level.
Manhattan chart of SNP markers significantly related to swine birth weight trait, as shown in FIG. 1, the ordinate is P value calculated by association analysis, i.e., -log10(pvTrue), the abscissa indicates the chromosome number, and the arrow indicates the 112031589 th nucleotide position on chromosome 4.
Second, multi-factor variance analysis of pig birth weight related SNP sites
The obtained SNP site g.112031589G/A, the effect of different genotypes in the experimental group 669 sows on the piglet birth weight was analyzed in this example. 674 sows were actually sequenced and mutated in this example, however, 5 individuals were removed by filtration during phenotypic quality control, and only 669 sows were used for subsequent analysis. The effect of different genotypes at the g.112031589G/A nucleotide sites on piglet birth weight was analyzed by multifactorial analysis of variance using the R software aov () function. The analytical model is as follows:
yijk＝u+Bi+HYSj+Mk+eijk
wherein, yijkFor newborn weight of piglets, BiEffect for the ith sow breed; HYSjThe-year-season effect for the jth birthing house; mkThe kth genotype at the g.112031589G/A locus; e.g. of the typeijkIs a random residual.
The study shows that the genotypes of g.112031589G/A comprise three types of GG, GA and AA, and the influence of different genotypes on the newborn weight of piglets is statistically analyzed, and the result is shown in the table 1. The results of the significance analysis for the three genotypes are shown in table 2.
TABLE 1 influence of different genotypes of g.112031589G/A on piglet birth weight
| Genotype(s) | Number of individuals | Mean birth weight (mean ± s.e.) |
| GG | 97 | 1.27±0.28kg |
| GA | 260 | 1.33±0.27kg |
| AA | 312 | 1.40±0.26kg |
TABLE 2P-value test results
| Genotype(s) | P-value |
| GG-AA | 0.0000085 |
| GG-GA | 0.0658851 |
| AA-GA | 0.0022926 |
The results in tables 1 and 2 show that in the experimental sow population, the birth weight of sows with the AA genotype at the g.112031589G/A site is significantly higher than that of sows with the GG genotype (P < 0.01); the g.112031589G/A locus has the birth weight of the sow with the AA genotype, which is higher than that of the sow with the GG genotype or the AG genotype (P is more than 0.05). Therefore, the A allele is the dominant allele related to the piglet birth weight, and the higher the number of the alleles, the higher the birth weight of the sow, or the trend is.
Third, establishment of birth weight related SNP locus detection method
1. Design of detection primers
In this example, for the obtained SNP site g.112031589G/A, upstream and downstream primers for detecting the site are designed for PCR detection of the SNP site. The upstream primer for detecting the SNP site g.112031589G/A is shown as Seq ID No.2, and the downstream primer is shown as Seq ID No. 3;
Seq ID No.2：5’-TCTTCGGGTCAGAAAGAGA-3’
Seq ID No.3：5’-CTTCCTGCAGATATTGGGG-3’。
2. extraction of pig genomic DNA
According to the whole genome DNA sequencing result, selecting a plurality of sow ear samples with GG, GA and AA genotype g.112031589G/A nucleotide sites, respectively, placing the selected sow ear samples in a centrifugal tube filled with 70% alcohol, and storing the sow ear samples in a refrigerator at-20 ℃ for later use. Extracting the genome DNA of the ear tissue by using the genome DNA extraction and quality detection method in the acquisition of SNP sites related to the birth weight of the first pig and the pig, detecting the quality and the concentration, and storing at the temperature of minus 20 ℃ for later use.
3. PCR amplification and sequencing of target fragment
PCR amplification is carried out using the extracted DNA as a template and based on the designed primers. The PCR amplification reaction system is as follows: 2.5 mu L of DNA template, 25 mu L of each 1.25 mu L, PCRMix reagent of 10mmol/L upstream primer and downstream primer, and 20 mu L of double distilled water; a total of 50. mu.L.
The PCR amplification conditions were: pre-denaturation at 98 ℃ for 2 min; then 35 cycles were entered: denaturation at 98 ℃ for 15s, annealing at 52 ℃ for 30s, and extension at 72 ℃ for 30 s; after the circulation is finished, the extension is carried out for 5min at 72 ℃ and the standby is carried out at 4 ℃.
A portion of the PCR amplification products was detected by electrophoresis on 1.5% agarose gel, and the results are shown in FIG. 2. In FIG. 2, the first lane is DNAmarker I, the second lane is the PCR amplification result of the genomic DNA of a sow of AA genotype, the third lane is the PCR amplification result of the genomic DNA of a sow of AG genotype, and the fourth lane is the PCR amplification result of the genomic DNA of a sow of GG genotype. The results in FIG. 2 show that the primers of this example can amplify a target fragment of about 244bp for all sows of different genotypes.
And (3) sending the residual PCR amplification product to Liuhua Dageno science and technology limited for sequencing, comparing and analyzing the sequencing result with the sequence of the related gene fragment of the pig in GenBank by using SnapGene software, and judging the genotype of g.112031589G/A. The sequencing sequence conforms to the sequence information described in SEQ ID NO.1, a SNP site g.112031589G/A exists at the 112031589 th base of the sequence, and partial sequencing results are shown in a figure 3.
SEQ ID NO.1：
5’-TCTTCGGGTCAGAAAGAGAGCACCTCCCCAGCCATCCCGGGGTTTTACAGGAATAAAGCTGCGTCATGTTCCCCGTCAGTTCAGGATTAAGAAATCTGATGAGTGTGGTTTTGCTGTTGATTGATGRGACATGCACGTTTGTGACTTCACGGCATGCATGGTTTAACATTCTCTTGTGCACCTTTACAGGCTGCCTTTGTTTATATTTTTGCCCAGATGGGTATCCCCCAATATCTGCAGGAAG-3’。
FIG. 3 shows the sequencing results for the AA genotype, AG genotype and GG genotype, respectively. The sequencing result shows that the SNP marker of the embodiment is located at 127 th base from the 5' end of the sequence shown in SEQ ID No.1, and the genotypes comprise three genotypes of AA, AG and GG.
The foregoing is a more detailed description of the present application in connection with specific embodiments thereof, and it is not intended that the present application be limited to the specific embodiments thereof. It will be apparent to those skilled in the art from this disclosure that many more simple derivations or substitutions can be made without departing from the spirit of the disclosure.
SEQUENCE LISTING
<110> Shenzhen Huashengshengsciences institute
SHENZHEN HUADA AGRICULTURAL APPLICATION Research Institute
<120> SNP marker related to birth weight of pig and application thereof
<130> 19I29274
<160> 3
<170> PatentIn version 3.3
<210> 1
<211> 244
<212> DNA
<213> 112031589 th nucleotide site amplification sequence on chromosome 4
<400> 1
tcttcgggtc agaaagagag cacctcccca gccatcccgg ggttttacag gaataaagct 60
gcgtcatgtt ccccgtcagt tcaggattaa gaaatctgat gagtgtggtt ttgctgttga 120
ttgatgrgac atgcacgttt gtgacttcac ggcatgcatg gtttaacatt ctcttgtgca 180
cctttacagg ctgcctttgt ttatattttt gcccagatgg gtatccccca atatctgcag 240
gaag 244
<210> 2
<211> 19
<212> DNA
<213> Artificial sequence
<400> 2
<210> 3
<211> 19
<212> DNA
<213> Artificial sequence
<400> 3
Claims (10)
1. An SNP marker associated with the birth weight trait of pigs, characterized in that: the SNP marker is located at 112031589 th nucleotide site on chromosome 4 of a reference sequence of version 11.1 of the porcine international genome and has G or A polymorphism.
2. The SNP marker according to claim 1, characterized in that: the SNP marker is located at the 127 th base from the 5' end of the sequence shown in SEQ ID No. 1.
3. The SNP marker according to claim 1 or 2, characterized in that: the SNP marker is that the birth weight of the piglet produced by the sow with the AA genotype is obviously larger than that of the sow with the AG genotype or the GG genotype.
4. A primer set for detecting the SNP marker according to any one of claims 1 to 3, wherein: the upstream primer of the primer pair is a sequence shown by Seq ID No.2, and the downstream primer is a sequence shown by Seq ID No. 3;
Seq ID No.2：5’-TCTTCGGGTCAGAAAGAGA-3’
Seq ID No.3：5’-CTTCCTGCAGATATTGGGG-3’。
5. a kit for detecting the SNP markers set forth in any one of claims 1-3, comprising: the kit comprises the primer pair of claim 4.
6. The kit of claim 5, wherein: the kit also comprises reagents for PCR amplification.
7. A method for detecting the SNP marker according to any one of claims 1 to 3, wherein: comprises the steps of carrying out PCR amplification on the genomic DNA of a sow to be detected by adopting the primer pair as claimed in claim 4 or the kit as claimed in claim 5 or 6, and obtaining the base condition of the SNP marker by determining the sequence of a PCR amplification product.
8. A method of predicting piglet birth weight in a piglet, comprising: comprises the steps of carrying out PCR amplification on the genomic DNA of a sow by using the primer pair as claimed in claim 4 or the kit as claimed in claim 5 or 6, obtaining the base condition of the SNP marker as claimed in any one of claims 1 to 3 by determining the sequence of the PCR amplification product, and predicting the pig birth weight of piglets born by the sow according to the base condition of the SNP marker.
9. Use of the SNP marker according to any one of claims 1 to 3 for prediction of porcine birth weight or for molecular assisted selection breeding of swine.
10. Use of the primer pair according to claim 4 or the kit according to claim 5 or 6 for pig birth weight prediction or molecular assisted selection breeding of pigs.
Priority Applications (1)
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