New molecular marker in MMP3 gene, used for DNA source tracing of pork products
A technology of molecular markers and genes, applied in the direction of DNA/RNA fragments, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of label loss, blurred label pattern, easy label replacement, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0017] Molecular marker search
[0018] (1) Primer design
[0019] In the NCB database, using the mRNA sequence of the pig MMP3 gene (NM_001166308) as a template, Primer 5.0 was used to design amplification primers for finding new SNP sites.
[0020] Forward primer: 5`-GCAGAAGTTCCTTGGGTTGG-3`
[0021] Reverse primer: 5`-AAACTTTTCCAGGTCCGTCAA-3`
[0022] The total volume of the PCR reaction is 20μl, including about 100ng of porcine genomic DNA, containing 1×buffer (Promega), 1.5mmol / L MgCl 2 , the final concentration of dNTP is 150 μmol / L, the final concentration of primer is 0.2 μmol / L, 2U Taq DNA polymerase (Promega). The PCR amplification program was: 94°C for 4min, then cycled 35 times (94°C for 30s, annealing at 56°C for 30s, 72°C for 20s), and finally 72°C for 10min. PCR reaction products were detected by 1.0wt% agarose gel electrophoresis.
[0023] (2) Cloning and sequencing analysis: the obtained MMP3 gene PCR product is cloned according to the following method:
...
Embodiment 2
[0031] Distribution of alleles
[0032] (1) Design of test groups:
[0033] Experimental group: samples of Meishan pigs (33 heads), Ningxiang pigs (24 heads), Pudong White pigs (24 heads), Duroc (10 heads), American Large White (34 heads) and French Large White (65 heads) were collected. Ear tissue, DNA extraction, a total of 190 DNA samples. The purpose of the test group is to detect the distribution of SNP molecular markers in different varieties.
[0034] (2) Genotype detection: use forward primer and reverse primer to amplify and detect all individual genotypes of the test population by HpyF3I-RFLP.
[0035] (3) Statistical analysis: record the genotypes of all individuals, and calculate the allele frequency, see Table 1 for the results.
[0036] Table 1 Distribution of PCR-RFLP-HpyF3I polymorphism in 6 pig breeds or strains
[0037]
[0038]Analyzing the distribution of the alleles of the SNP site, it was found that the allele frequency of the molecular marker in d...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 