Gene clone and expression of RGD die body protein of oral gland in Japan lamprey possessing function for anti tumour
A technology of oral gland and lamprey, applied in the field of gene cloning and expression of RGD motif protein of Japanese lamprey oral gland with anti-tumor effect
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[0062] 1. Extraction of total RNA: Trizol reagent of GIBCOBRL was used. details as follows:
[0063] (1) Get the venom glands of Agkistrodon halys and quickly place them in liquid nitrogen for preservation;
[0064] (2) Weigh 0.2 g venom gland tissue, add 1 ml TRIzol reagent to prepare venom gland homogenate, and incubate at 4°C for 5 min;
[0065] (3) Add 0.2ml chloroform, close the lid tightly, shake vigorously for 15sec, and then place it on ice for 5min;
[0066] (4) 4°C, 12000xg, centrifuge for 15 minutes;
[0067] (5) Transfer the upper aqueous phase to another centrifuge tube, add 0.5ml of isopropanol, and incubate on ice for 10 minutes;
[0068] (6) 4°C, 12000xg, centrifuge for 15 minutes;
[0069] (7) Discard the supernatant, add 1ml of 75% ethanol to the precipitate (containing RNA) to wash, and vortex to mix;
[0070] (8) Centrifuge at 10,000xg for 5 minutes at 4°C to obtain RNA precipitates;
[0071] (9) After air drying, dissolve with appropriate amount of T...
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