SNP molecular marker related to body size traits of Macrobrachium rosenbergii and application thereof
A technique of Macrobrachium rosenbergii and molecular markers, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, DNA/RNA fragments, etc., to achieve the effects of accelerating genetic progress, saving production costs, large economic application value and scientific research value
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[0025] 1. Detection of SNP molecular markers
[0026] 1. Designing primers: designing a pair of primers for specifically amplifying the gene fragment containing the T574C site of the Macrobrachium rosenbergii ND1 gene.
[0027] Utilize the software Primer 5.0, use the ND1 gene (GenBank: NC_006880.1) of Macrobrachium rosenbergii in GenBank as the specificity amplification primer of reference design, the primer sequence that amplifies is
[0028] Upstream F: 5'-GATAAAGTAGGCTATGCTGG-3' (SEQ ID №1), 20bp;
[0029] Downstream R: 3'-CCTCCTCTGTACTCGGTATT-5' (SEQ ID No. 2), 20 bp.
[0030] The optimum temperature is 48.8°C, the product length is 542bp, and it was synthesized by Guangzhou Sangon Bioengineering Co., Ltd.
[0031] 2. Extraction of Macrobrachium rosenbergii Genomic DNA
[0032] Using OMEGAD whole blood kit, the specific steps are as follows:
[0033] (1) Take 150 μL blood sample for each sample, add Elution Buffer, and vortex for 10 seconds;
[0034] (2) Add 500 μL o...
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