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Ensure that all of the purification buffers are at four °C and carry out the protein purification within the chilly room to avoid protein denaturation. BE AWARE: After the coupling of the first C-terminal amino acid of all peptides to the amino group of the membrane, the amino teams between the spots and also among the amino teams inside the spot areas don't kind bonds with the amino acids. NOTE: This avoids the coupling of amino acids within the further cycles with the membrane instead of the growing peptide chain.
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