Novel method for obtaining goat-induced multi-potent stem cells
A technology of pluripotent stem cells and a new method, which is applied in the fields of stem cell engineering, preservation of rare species, and tissue and cell engineering, can solve the problems of long production cycle, low safety and cumbersome operation of goat iPS cells, and achieve production efficiency and safety The effect of high safety, high safety and high production efficiency
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[0018] 1. Cloning of pluripotency transcription factors
[0019] ①Total RNA extraction
[0020] The total RNA of goat testis tissue, skin tissue and small intestine tissue was extracted respectively by RNA extraction kit, and the extraction steps were as follows:
[0021] (1) Add 50-100mg of goat testicular tissue, skin tissue and small intestine tissue samples to the mortar treated with DEPC water, add liquid nitrogen and quickly grind to powder, put them into 1.5mL EP tubes, and quickly Add 1mL of Trizol, mix well and let stand at room temperature for 10 minutes;
[0022] (2) Centrifuge in a refrigerated centrifuge at 4°C and 12,000 rpm for 10 minutes, transfer the supernatant to a new EP tube, add 200 μL of chloroform and shake vigorously repeatedly for 15 seconds, place at 25°C for 3 minutes, and then centrifuge for 15 minutes with the same standard;
[0023] (3) Transfer the upper aqueous phase in the EP tube to another new tube, add 400 μL of isopropanol, mix well, lea...
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