Alkali-tolerant SNP marker for Mozambican tilapia and its application
A tilapia, alkali-resistant technology, applied in climate change adaptation, recombinant DNA technology, DNA/RNA fragments, etc., to achieve the effect of great scientific value and commercial value
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Embodiment 1
[0039] Research on the role of SNP markers in this example:
[0040] The Mozambican tilapia (Oreochromis mossambicus) (500 tails) used in this example were all from the Gaoyao Aquatic Germplasm Base, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences. The alkalinity 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 sodium bicarbonate (NaHCO) every 20 minutes. 3 ) solution to increase the carbonate alkalinity of the pool water, increase the alkalinity by 1g / L each time, measure the actual carbonate alkalinity of the water body by acid-base titration, titrate with 1mol / L hydrochloric acid solution (HCl), and use methyl orange Check the actual carbon...
Embodiment 2
[0046] In this example, the SNP markers of Mozambique tilapia are screened / detected:
[0047] (1) Extraction of sample DNA
[0048] (1) Take 3 mg of the fin ray tissue of Mozambique tilapia as a sample to be tested, cut into pieces, and add 0.5 mL of lysis solution (10 mmol / L Tris-HCl; 0.1 mol / L EDTA; 0.5% SDS; 30 mg / L RNase; 100 mg / L proteinase K, pH 8.0), digested at 55°C for 1 hour;
[0049] (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;
[0050] (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;
[0051] (4) Wash the precipitate once wi...
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