A SNP marker related to salt tolerance of tilapia and its application
A tilapia and salt-tolerant technology, which can be applied in climate change adaptation, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of low selection rate and long cycle, and achieve good specificity, great scientific value and commercial value. The effect of high value and amplification efficiency
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Embodiment 1
[0037] Research on the effect of the SNP marker of the present invention:
[0038]The Oreochromis hornorum (500 tails) used in the present invention all come from the Gaoyao Aquatic Germplasm Base of the Pearl River Fisheries Research Institute of the Chinese Academy of Fishery Sciences. The salinity stress experiment was carried out in a fish pond of 320cm×200cm×150cm. Before the experiment, the cells were incubated for 7 days, during which the dissolved oxygen was 8.7±0.6mg / L, the water temperature was 28.5±1℃, and the pH value was 8.4±0.5. Select healthy, harmless and lively tilapia for the experiment. After the experiment starts, add pre-prepared salt water of about 113‰ every 30 minutes to increase the salinity of the pool water, and increase the salinity by 5‰ each time. Use a hand-held salinity meter Check the actual salinity of the pool water. During the period of continuous aeration, no feed was fed. After 2 hours, tilapia began to die, and the death time, water sa...
Embodiment 2
[0044] The present embodiment screens / detects the SNP markers of tilapia:
[0045] (1) Extraction of sample DNA
[0046] (1) Take 3 mg of tilapia fin ray tissue of the sample to be tested, cut into pieces, add 0.5 mL of lysis solution (10 mmol / LTris-HCl; 0.1 mol / L EDTA; 0.5% SDS; 30 mg / L RNase; 100 mg / L) L proteinase K, pH 8.0), digested at 55°C for 1 hour;
[0047] (2) Add an equal volume of phenol / chloroform / isoamyl alcohol (volume ratio 25:24:1) mixture, mix well, stand at room temperature for 5 minutes, centrifuge at 12,000 rpm for 10 minutes, take the supernatant, and then Extract once with chloroform, stand at room temperature for 5 minutes, centrifuge at 12,000 rpm for 10 minutes, and take the supernatant;
[0048] (3) adding 2 times the volume of absolute ethanol, standing at room temperature for 10 minutes to precipitate DNA, and centrifuging at 12,000 rpm for 10 minutes to obtain a precipitate;
[0049] (4) Wash the precipitate once with 70% ethanol, centrifuge at...
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