AAT Bioquest

How do I prepare a spin column?

Posted April 2, 2019

I'd like to use a 1 mL spin column, such as a Sephadex® G-25, BioGel® P-6 DG Media or other desalting column to purify 100 µL of a reaction mixture. Can you provide a sample protocol?


Below is a sample procedure on how to set up and use a 1 mL spin column for purification of 100 µL test conjugate:

  1. Fill with beads
  2. Drain the solution, top dry
  3. Buffer exchange using ~4 mL PBS buffer (pH 7.2 - 7.4)
  4. Spin column at 2000X for 2 minutes
  5. Exchange current 15 mL tube for a brand new one
  6. Load sample mixture directly to the top of the desalting column
  7. Add 10 µL of PBS buffer (pH 7.2 - 7.4)
  8. Spin column at 2000X for 5 minutes
  9. Collect elution
  10. Measure the concentration using a spectrophotomer
Additional resources

Microarray Methods and Protocols