Method for identifying Bama minipigs through ISSR molecular marking
A Bamaxiang pig and molecular marker technology, applied in biochemical equipment and methods, microbiological measurement/inspection, etc., can solve problems such as inability to judge unknown samples, achieve obvious differences, reduce identification errors, and ensure varieties Effect
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Embodiment 1
[0025] A method of UPGMA cluster analysis using ISSR molecular markers to identify Bamaxiang pigs.
[0026] The method for identifying Bama fragrant pigs comprises the following steps:
[0027] A. Sample DNA extraction;
[0028] B. PCR reaction and product detection;
[0029] C. UPGMA cluster analysis, if the similarity coefficient with the standard product is ≥0.94, the sample can be identified as Bamaxiang pig.
[0030] The nucleotide sequence of the primer used in the PCR reaction is 5'-(ATG) 6 -3'.
Embodiment 2
[0032] Bama pig, Huanjiang pig, Tibetan pig, Bama native pig and Chenghua pig were identified by ISSR molecular markers.
[0033] Weigh 0.5 g of fresh pork samples to be identified from Bama Xiang pig, Huanjiang Xiang pig, Tibetan pig, Bama local pig and Chenghua pig, grind with liquid nitrogen, and extract DNA according to the instructions of the DNA extraction kit. The DNA extraction kit is Tiangen blood / cell / tissue genomic DNA extraction kit TIANampGenomicDNAKit (DP304-02). Use 0.8% agarose gel electrophoresis to detect the integrity of the extracted DNA, and use an ultra-micro protein nucleic acid analyzer to detect the mentioned DNA quality and OD 260nm and OD 280nm Concentration, determination of DNA purity OD 260nm / OD 280nm . Store at -20°C for later use.
[0034] PCR reaction primer, its nucleotide sequence is 5'-(ATG) 6 -3'. The ISSR was amplified using the primers. The PCR reaction system is: template DNA 1.5 μL, primer 1 μL, 2×TaqPCR MasterMix 12.5 μL, ddH ...
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