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Nutrients, 12 (2), 334

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Embodiment 3 [0053] Expression and purification of embodiment 3 fusion protein [0054] Transfer the seed liquid of the recombinant positive engineering strain of Example 2 to LB medium according to the inoculum size of 2%, cultivate it at 37C until OD600=0.6, add 50 l of 0.5mol / L IPTG, induce at 15C for 16 hours, and collect by centrifugation For each 1 g of wet bacteria, resuspend in 10 mL of lysis buffer (50 mM Tris-HCl, 50 mM NaCl, 5% glycerol, pH 8.0, 17 l (10 mg / mL PMSF)), ultrasonicate and centrifuge to obtain supernatant and precipitate

You are not becoming "frail"

Here are some of the obvious things you may notice after switching