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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 some of the amino groups inside the spot areas do not type bonds with the amino acids. OBSERVE: This avoids the coupling of amino acids in the further cycles with the membrane as a substitute of the growing peptide chain. Ensure all the purification buffers are at 4 °C and carry out the protein purification in the cold room to keep away from protein denaturation.

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