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Never freeze reconstituted peptide, as ice crystal formation destroys the molecular structure

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The preparation method according to claim 1 or 2, wherein the coupling in step 2 is performed by condensing by DIC+A or B+A+C method until the resin is detected to be transparent with ninhydrin, wherein A is chosen from HOBt or HOAt, B is chosen from HBTU, HATU, TBTU, PyAOP, or PyBOP, C is chosen from DIPEA or TMP