FAQ: Should the crude extract be diluted with lysis buffer to increase the volume for the binding step?

Dilution of the crude extract may be performed if you are unable to ensure efficient mixing of the beads with the low volume of extract. Dilution of crude extract may result in a slight decrease in yield. In addition, the binding time may be extended up to 1 hour to maximize yield. Another option is to use a sample mixer that provides a vibration function (e.g., HulaMixer, Thermo Catalog #15920D) which is able to agitate the beads and keep them thoroughly re-suspended in small volumes.