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WORD: This avoids the coupling of amino acids within the further cycles with the membrane as an alternative of the growing peptide chain. Be certain that all of the purification buffers are at four °C and carry out the protein purification in the chilly room to avoid protein denaturation. BE AWARE: After the coupling of the primary C-terminal amino acid of all peptides to the amino group of the membrane, the amino teams between the spots and in addition some of the amino teams inside the spot areas don't form bonds with the amino acids.