Reagent Preparation: Theoretical and Practical Discussions

Deb N. Chakravarti1, Bulbul Chakravarti1, Buddhadeb Mallik2

1 York College/CUNY, Jamaica, New York, 2 Montgomery College, Department of Chemistry, Rockville, Maryland
Publication Name:  Current Protocols Essential Laboratory Techniques
Unit Number:  Unit 3.1
DOI:  10.1002/9780470089941.et0301s9
Online Posting Date:  October, 2014
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Abstract

One of the most important requirements for success with any biochemical protocol is the accurate preparation of reagents. This unit provides general guidelines for the preparation of reagents, including 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, and avoidance of chemical or microbial contamination. 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. © 2014 by John Wiley & Sons, Inc.

Keywords: buffer; solution; reagent; molarity; normality; concentration

     
 
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Table of Contents

  • Reagent Preparation
  • Accuracy of Weighing and Pipetting
  • Use of Calibrated pH Meters
  • Avoiding Chemical and Microbial Contamination of Reagents
  • Preparing Reagent or Buffer Solutions
  • Making Buffer Solutions
  • Literature Cited
     
 
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Materials

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Literature Cited

Literature Cited
  Blanchard, J.S. 1984. Buffers for enzymes. Methods Enzymol. 104:404‐414.
  Fernbach, A. and Hubert, L. 1900. De l'influence 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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