Molecular marker identification method related to pig birth litter weight and fetal weight and application of molecular marker identification method
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A technology of molecular markers and identification methods, applied in the fields of biochemical equipment and methods, microbial determination/inspection, recombinant DNA technology, etc.
Active Publication Date: 2016-10-12
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI +1
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[0007] At present, there are few reports on the function of the pig CYP19A1 gene. The applicant used the method of genome-wide association analysis to conduct an association analysis be
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Embodiment 1
[0092] A molecular marker identification method related to pig birth weight and fetal weight:
[0093] (1) Sample collection: put in 75% alcohol and store at -20°C, DNA to be extracted;
[0094] (2) Extraction and detection of pig genomic DNA:
[0095] 1) Genomic total DNA was extracted from pig tissue;
[0096] 2) After the DNA is fully dissolved, measure the concentration of the extracted DNA with a UV spectrophotometer, and detect the quality of the extracted DNA by agarose gel electrophoresis;
[0097] (3) SNP chip genotype determination and quality control of genotype data:
[0098] 1) Take 2ul stock solution for electrophoresis detection
[0099] 2) Primer design: According to the SNP site sequence information, use the primer design software Assaydesign3.1 of sequenom company to design and synthesize the PCR reaction and single-base extension primers;
[0100]
[0101] Take 2.5ul each of Forward PCR primer and Rewerse PCR primer for each SNP, and mix them into a 5...
Embodiment 2
[0113] A molecular marker identification method related to pig birth weight and fetal weight:
[0114] 1. Primer design:
[0115] 1st-PCRP: forward primer (F)
[0116] 2nd-PCRP: reverse primer (R)
[0117] UEP_SEQ: Single base extension primer
[0118] UEP_MASS: single base extension primer quality
[0119] UEP_DIR: the DNA strand where the single base extension primer is located
[0120] F: chain of justice
[0121] R: antisense strand
[0122] 2. Experimental process:
[0123] a. First dilute the primer well, primer dilution:
[0124] (1) For each SNP, there are three primers, and the numbers of these three primers are marked on the cap of the primer synthesis tube.
[0125] (2) Forward PCR primers and Rewerse PCR primers are first centrifuged and then added with water. The amount of water added is related to the OD value (the primer tube and the primer design table are marked), and 36ul of water is added for every 1OD. After adding water, let stand at room temperat...
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Abstract
The invention discloses a molecular marker identification method related to pig birth litter weight and fetal weight and an application of the molecular marker identification method. The identification method comprises the following steps: collecting a sample; extracting and detecting pig genome DNA; then, conducting SNP (single nucleotide polymorphism) chip genotype judgment and genotype data quality control: adding a PCR amplification primer pair and conducting a PCR amplification reaction by taking DNA of the sample as a template, purifying a reaction product and co-crystallizing the purified reaction product with a mass spectrum chip, conducting detection and typing by virtue of a flight mass spectrum method, detecting an SNP site of a target gene, computing an SNP genotype and an allele frequency by virtue of PopGene3.2, and conducting association analysis on the SNP and characters by virtue of SAS9.2 software; and finally, conducting data collating and analysis. The identification method disclosed by the invention, by determining the genotype of a pig individual, can assist identification of multiparous litter weight and fetal weight.
Description
technical field [0001] The invention belongs to the technical field of breeding pig production research, and relates to the research on SNP molecular markers of farrowing traits of pig genes, in particular to an identification method and application of molecular markers related to newborn litter weight and fetal weight of pigs. Background technique [0002] my country's pig breeding industry has a long history, and the pig breed resources are abundant and widely distributed, with huge utilization potential and broad development prospects. The production of breeding pigs is the foundation of the pig industry. Only through scientific feeding and management can the breeding pigs breed a large number of high-quality piglets and improve the economic benefits of the pig industry. [0003] With the continuous and rapid development of my country's economy, people's living standards are gradually improving, and the demand for pork is also increasing. Therefore, people are also paying...
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