Tumor necrosis factor of Chinese mitten crab and its application
A tumor necrosis factor, Chinese mitten crab technology, applied in the direction of tumor necrosis factor, application, cytokine/lymphokine/interferon, etc., can solve problems such as economic losses in the crab breeding industry
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[0036] 1. Extraction of total RNA from Eriocheir sinensis
[0037] Extract healthy Chinese mitten crabs (about 40g each, purchased at the mouth of the Chinese mitten crab breeding pond in Baoying County, Yangzhou City, Jiangsu Province, collected by Huang Ying, collection date 2018.06.18) blood cells, heart, hepatopancreas, gills, muscles , gut, nerve and eye stalk tissue. In addition, 50 μL of Staphylococcus aureus and Vibrio parahaemolyticus (about 3×10 7Bacteria) were injected into the second step of the river crab, and the hepatopancreas tissues of different time periods (2, 6, 12, and 24 h) were extracted to analyze the changes of EsTNF expression after pathogenic bacteria stimulation.
[0038] (1) Add 1 mL of TRIzol reagent to 100 mg of tissue, fully homogenize in an ice bath; then transfer to a 1.5 mL centrifuge tube, and let stand at 15-30°C for 5 minutes;
[0039] (2) Add 200 μL of chloroform, shake vigorously for 30 seconds and let stand at 15-30°C for 5 minutes; ...
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