By Glenn Sant
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Extra info for Marine Invertebrates of the South Pacific: An Examination of the Trade (Species in Danger)
Example text
Methods (see Notes 1–11) 1. Take the glass plates, spacers, and comb appropriate to the gel apparatus to be used. Thoroughly clean them by washing in soapy water, rinse in distilled water, and then methanol. Allow to air-dry. Assemble plates and spacers as instructed by suppliers in preparation for making the gel. 2. Prepare the separating gel mixture. 1 cm. For gel(s) of 15% T, the mixture is made as follows. 5 mL of separating gel buffer A, 45 µL stock ammonium persulphate solution and 15 µL of TEMED.
7. 8. 9. 10. 25 mL of stock separating gel buffer A, 15 µL stock ammonium persulphate, and 5µL TEMED. Mix gently and use immediately. Pour off the butanol from the polymerised separating gel. Rinse the top of the gel with a little water, then a little upper gel mixture (from step 4). Fill the gap remaining above the gel with the upper gel mixture from step 4 Insert the wellforming comb without trapping any air bubbles. 5 h). When the gel has finally polymerized store it at 4°C overnight or longer (see Note 9).
Wilkins, M. R. and Gooley, A. (1997) Protein identification in proteome analysis, in Proteome Research: New Frontiers in Functional Genomics (Wilkins, M. , Williams, K. , Appel, R. D. and Hochstrasser, D. ), Springer-Verlag, Berlin, pp. 35–64. 15. Patterson, S. , and Goodlett, D. R. (2001) Mass spectrometrybased methods for protein identification and phosphorylation site analysis, in Proteomics, From Protein Sequence to Function (Pennington, S. R. and Dunn, M. ), BIOS Scientific Publishers, Oxford, pp.