Team:Freiburg/Protocols/Desalting

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Desalting using BioRad desalting columns

material: Lysis buffer, aqua dest, 20% EtOH
duration: 1h

equlibration

  • put columns on a falcon labeled 'waste'
  • clean columns from EtOH by washing 4-5 times with 5 mL of aqua dest
  • equilibrate with the same amount of lysis buffer
    • let buffer flush into the column and close just before column starts to run dry

loading

  • add 2.5 mL of protein solution
    • let again just flow into the column
  • Change the falcon to a new one in which the actual elution should take place
  • repeat the following step 3 times
    • pour 2mL of lysis buffer on the column and collect the flow-through (just when solution is flushed in)

cleaning

  • clean the column by washing 4-5 times with aqua dest for storage or lysis buffer for further use
  • for storage: equilibrate the column with 2-3 times with 20% EtOH solution and store @ 4°C

Original protocol

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