Column Chromatography

C.L.F. Meyers1

1 Purdue University, West Lafayette, Indiana
Publication Name:  Current Protocols in Nucleic Acid Chemistry
Unit Number:  Appendix 3E
DOI:  10.1002/0471142700.nca03es03
Online Posting Date:  May, 2001
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

Column chromatography is used extensively in nucleic acid chemistry to purify or characterize products from chemical reactions. This appendix focuses on separations using silica gel or alumina. The procedures described include loading the column, assembling the apparatus, separating compounds and collecting fractions, and optimizing chromatography conditions.

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Basic Protocol 1: Column Chromatography
  • Alternate Protocol 1: Microscale Chromatography
  • Support Protocol 1: Packing a Chromatography Column
  • Literature Cited
  • Figures
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

Basic Protocol 1: Column Chromatography

  Materials
  • Crude mixture
  • Eluent
  • Chromatography column, packed (see protocol 3)
  • Flash chromatography apparatus (optional): 250 mL solvent reservoir, 35/20 spherical joint with appropriate flow controller (Chemglass), and air source
  • Additional reagents and equipment for thin‐layer chromatography (TLC; appendix 3D)

Alternate Protocol 1: Microscale Chromatography

  Materials
  • Eluent
  • Adsorbent (e.g., mesh silica gel; Fisher)
  • Glass wool
  • Glass column with ground glass or threaded joint at the top and stopcock at the bottom
  • Fine sand
  • Stiff rubber tube or rubber stopper attached to a handle (e.g., wooden or glass rod)
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

Literature Cited
   Mayo, D.W., Pike, R.M., and Trumper, P.K. 1994. Microscale Organic Laboratory: With Multistep and Multiscale Syntheses, 3rd ed. John Wiley & Sons, New York.
   Still, C.W., Kahn, M., and Mitra, A. 1978. Rapid Chromatographic technique for preparative separations with moderate resolution. J. Org. Chem. 43:2923‐2925.
   Zweig, G. and Sherma, J. 1972. CRC Handbook of Chromatography, Volumes I and II. CRC Press, Cleveland, Ohio.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library