Alkali-resistant SNP (Single Nucleotide Polymorphism) marker for momulberry bike
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]Study on the action of SNP tag of this example:
[0040]The Mozambian Mossambicus (500 tails) used in this example comes from the high demanded aquatic breeding base of the Zhujiang Aquatic Research Institute, China Aquatic Science Research Institute. The alkali stress experiment is carried out in a fish pool of 320 cm × 200 cm × 150 cm. Relaying 7 days before the experiment, dissolved oxygen during 8.7 ± 0.6mg / L, water temperature is 28.5 ± 1 ℃, pH value 8.4 ± 0.5. Choose a healthy and hurtful, lively tilapia, after the experiment, add pre-pre-pre-pre-pre-pre-pre-predicted sodium bicarbonate every 20 minutes (Nahco3The solution increases the carbonate alkali of the pool water, and the alkalinity ratio of 1 g / L is increased each time, the actual carbonate degree of the water is measured with acid-base titration, titration with 1 mol / L hydrochloric acid solution (HCl), and methyl orange The actual carbonate level of the pool water is detected in the indicator. During the per...
Embodiment 2
[0046]This embodiment of the SNP markers Mozambique tilapia screening / detection:
[0047]Extraction (a) sample DNA
[0048](1) Retrieving the test sample Mozambique non-fish fin tissue 3 mg, after cutting, 0.5 ml of lysate (10 mmol / L Tris-HCl; 0.1 mol / L EDTA; 0.5% SDS; 30 mg / L RNase; 100 mg / L protease K, pH 8.0), digested at 55 ° C for 1 hour;
[0049](2) Add equal volume of phenol / chloroform (volume ratio 25: 24: 1) mixture, mix, after standing for 5 minutes, 12000 rpm, centrifuge for 10 minutes, take the supernatant, and then Take it once with chloroform, after standing for 5 minutes at room temperature, centrifugation 12000 rpm for 10 minutes, take the supernatant;
[0050](3) 2x volume of anhydrous ethanol was added, and it was allowed to stand for 10 minutes to precipitate DNA, and 12000 rpm was centrifuged for 10 minutes to obtain a precipitate;
[0051](4) The precipitate was washed once with 70% ethanol, 12000 rpm for 2 minutes, suction of the supernatant, and allowed to stand ...
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