A kind of lymphokine tal and its coding gene and application
A technology of lymphokines and genes, applied in the direction of cytokines/lymphokines/interferons, applications, genetic engineering, etc., can solve problems that restrict the healthy and sustainable development of oyster farming in Hong Kong, the death of Hong Kong oysters, and the destruction of the breeding environment, and achieve The effect of promoting the proliferation of blood lymphocytes
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Embodiment 1
[0018] 1. RNA extraction
[0019] Hong Kong oyster total RNA was extracted using Trizol (Invitorgen), and the specific steps were as follows: (1) Take 50 mg of Hong Kong oyster tissue in a liquid nitrogen pre-cooled mortar, grind the tissue into powder, and transfer it to a centrifuge containing 1 ml Trizol. (2) Add 200 μL of chloroform, shake vigorously for 40 seconds, and place at room temperature for 5 minutes; (3) Centrifuge at 12,000×g for 10 minutes at 4°C, transfer the supernatant to a new tube; (4) Add 0.5ml of Isopropanol, mix well, precipitate overnight at -80°C; (5) centrifuge at 4°C, 12,000×g for 10 min, discard the supernatant; (6) wash the precipitate twice with 75% ethanol; (7) discard the supernatant, add 50 μL DEPC Water dissolves RNA.
[0020] 2. cDNA library construction
[0021] Full-length cDNA library construction using GeneRacer TM cDNA library construction Kit (Invitrogen), in which the specific steps of 5'RACE are as follows: (1) Calf intestinal ph...
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