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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 likewise some of the [amino teams](http://git.gkcorp.com.vn16000/katlynchristma/6037solicitor-uk/wiki/TiE-Con-TiE-Con) within the spot areas don't kind bonds with the amino acids. Be certain that all the purification buffers are at 4 °C and carry out the protein purification in the chilly room to avoid protein denaturation. NOTICE: This avoids the coupling of amino acids in the additional cycles with the membrane as an alternative of the rising peptide chain.
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