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coli ., and the positive vectors were picked out from culture plates and amplified in LB liquid medium at 37 C, 250 rpm until the OD 600 reaching 0.81.0

For each strain, 200 mL of culture was harvested by centrifugation at 5,000 rpm for 15 min under aseptic conditions, and the resulting cell pellets were resuspended in fresh medium

Thats why subcutaneous injection of large doses (5-10 mg) is the standard delivery method in most epithalon protocols: you have to flood the system to get enough surviving peptide into target tissues before degradation wipes it out

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It is always recommended you obtain a research investigator (RI)