Reagent Preparation: Theoretical and Practical Discussions
1Proteomics Center, Keck Graduate Institute of Applied Life Sciences, Claremont, California
Abstract
One of the most important requirements for the successful completion of any biochemical protocol is to prepare reagents accurately. This unit describes general guidelines for the preparation of reagents, use of clean glass- and plasticware, use of high-purity reagents, safe handling of chemicals and biochemicals, use of high-purity distilled or deionized water, accurate weighing, pipetting, and pH measurement, and avoidance of chemical or microbial contamination of reagents and use of sterile reagents. Different units of concentration used for preparation of reagents or buffer solutions are described. This unit also includes a section for the preparation of buffer solutions with a practical example.
Keywords: biochemicals; buffer; chemicals; deionized water; distilled water; glassware; pH; plastic ware; reagents; units of concentration
Literature Cited
| Literature Cited | |
| Blanchard, J.S. 1984. Buffers for enzymes. Methods Enzymol. 104:404-414. | |
| Fernbach, A. and Hubert, L. 1900. De linfluence des phosphates et de quelques autres matières sur la diastase protèolitique du malt. Compt. Rend. 131:293-295. | |
| Gabler, R., Hegde, R., and Hughes, D. 1983. Degradation of high purity water on storage. J. Liq. Chromatogr. 6:2565-2570. | |
| Gomori, G. 1955. Preparation of buffers for use in enzyme studies. Methods Enzymol. 1:138-146. | |
| Good, N.E. and Izawa, S. 1972. Hydrogen ion buffers. Methods Enzymol. 24:53-68. | |
| Seed, B. 2003. Silanizing glassware. Curr. Protoc. Mol. Biol. 28:A.3B.1-A.3B.2. | |
| Stoll, V.S. and Blanchard, J.S. 1990. Buffers: Principles and practice. Methods Enzymol. 182:24-38. | |
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