How-To-Do-It A Rapid, Simple & Inexpensive Experiment in Gel Filtration Chromatography

We have developed a simple and inexpensive experiment using gel filtration chromatography that can be performed in less than 90 minutes. This procedure not only provides students with hands-on experience in both the preparation and use of chromatography columns, but also uses colored molecules to provide dramatic visual confirmation of the process of size-exclusion chromatography. Additionally, the column can be calibrated using molecules of known molecular weights and can then be used to estim a t e t he mo le cu la r w ei gh ts of unknown samples. Our gel filtration experiment costs only a fraction of that of commercially marketed laboratory exercises. A basic understanding of methods and techniques in modern life science research is important in understanding the way that life scientists develop new methods for answering biological questions and the potentials and limitations of these modern experimental methods. On both the introductory college and high school levels, many students are performing basic exercises using electrophoresis and recombinant DNA. Unfortunately, few of these students are exposed to chromatography, the major means of purifying proteins and other biological molecules. This is partly because of the expense and complexity of many chromatographic procedures. Our experiment involves a simplified version of the analytical methods used by research laboratories to frac-