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1. Each SPOT carries approximately 5 nmol of peptide, covalently bounded to the cellulose membrane.
2. The yield and purity of each peptide will vary depending on the sequence. There is a spacer of two ß-alanines between the peptide and the cellulose membrane.
3. The peptides are synthesized in a grid with a spacing of 0.37 cm x 0.37 cm (0.14 in x 0.14 in), 20 peptides in each row. SPOT orientation is read from left to right across the grid.
4. The beginning and the end of each of the rows are marked with pencil lines. The first peptide is marked with “1”.
5. A numbered sequence list and diagram of the SPOT array is provided for peptide sequence determination.
Please Note: Sometimes the SPOTs are difficult to visualize due to short peptide sequences and amino acid composition of the sequences. The SPOTs will typically become more visible during the incubation process. Alternatively, the membrane can be placed under UV light for visual detection.
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