View Single Post
Old 05-30-2018, 05:40 AM   #2
ATϟGC
Member
 
Location: Canada

Join Date: Jun 2013
Posts: 54
Default

I find it can be hard to get more than 80-90% recovery and I sometimes get as low as 50%.

Here are a few suggestions you can try for improving recovery:

-Do not dry the beads too long. If you see cracks forming in the bead pellet then they have been dried too long and elution of the DNA from the beads can be difficult. I try to dry the beads to the point they look like a wet or moist clay and then add the elution buffer. Over-drying the beads is a common cause of poor recovery.

-Elute at 50-60 degrees Celsius for 30 minutes or longer. You may also leave them at 4 degrees for a few days to help with elution.

-Try a custom elution buffer containing tween-20 such as: 10 mM tris-HCl pH 8.0, 0.1 mM EDTA pH 8.0, and 0.1-0.5% tween-20 v/v.

Please note that I have not rigorously tested these modifications in a systematic manner and they might not help significantly.


It would be helpful to know what size range of DNA you are trying to purify, what beads you are using and a description of your protocol.
ATϟGC is offline   Reply With Quote