Recombinant L-251 protein with anti-inflammatory effect secreted by Japanese lamprey oral gland
A technique for oral glands and lampreys, which is applied in the field of recombinant Japanese lamprey oral glands to secrete L-251 protein with anti-inflammatory effect, which can solve problems such as tissue and cell damage.
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[0050] 1. Extraction of total RNA: Trizol reagent from GIBCO BRL was used. details as follows:
[0051] (1) Take the Japanese lamprey oral gland and place it in liquid nitrogen to grind rapidly;
[0052] (2) Weigh 0.2 g glandular tissue and secretion, add 1 ml TRIZOL reagent to prepare homogenate, and incubate at 4°C for 5 min;
[0053] (3) Add 0.2ml chloroform, close the cap tightly, shake vigorously for 15 seconds, and then place on ice for 5 minutes;
[0054] (4) 4°C, 12000g, centrifuge for 15 minutes;
[0055] (5) Transfer the upper aqueous phase to another centrifuge tube, add 0.5ml isopropanol, and incubate on ice for 10 min;
[0056] (6) 4°C, 12000g, centrifuge for 15 minutes;
[0057] (7) Discard the supernatant, add 1ml of 75% ethanol to the precipitate (containing RNA) to wash, and vortex to mix;
[0058] (8) Centrifuge at 10,000 g for 5 min at 4°C to obtain RNA pellets;
[0059] (9) After air drying, dissolve with appropriate amount of TE or RNase-free water f...
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