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OBSERVE: 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 do not form bonds with the amino acids. NOTE: This avoids the coupling of amino acids in the further cycles with the membrane as an alternative of the growing peptide chain. Ensure that all the purification buffers are at four °C and carry out the protein purification in the cold room to keep away from protein denaturation.
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