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●●●●●　SEPHADEX G-50 SPIN COLUMN　●●●●●



  1. PART I (preparing the Sephadex)

    1. Dissolve 5g G-50 Sephadex ( medium) in 100ml 1X TE.
    2. Heat to 90 degrees. Decant off the buffer and refill. Repeat this three times.
    3. Store at 4 degrees under 1X TE: add Na azide to 0.05 % for long term storage.

  1. PART II (preparing the column)

    1. Plug the end of an Evergreen P-1000 pipet tip with a small amount of glass wool.
    2. Place the Evergreen tip in a Sarstedt # 55.480 tube.
    3. Add hydrated Sephadex G-50 to top of the P-1000 tip.
    4. Let this settle for appx. 5 min.
    5. Discard liquid in bottom of tube.
    6. Spin the column 6K, 75 seconds in the table top centrifuge.
    7. Discard tube and replace ; load your sample.
    8. Spin the column 6K, 75 seconds.
    9. Load an additional 50 ul 1X TE on the column.
    10. Spin the column for another 75 seconds at 6K.
    11. Collect your sample.

      *** Do not assume that this protocol will remove all of the low molecular weight material form your prep.***

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