[Erik]

I’m using this protocol to clean up DNA template after a linearization reaction (removal of buffer & enzyme, rather than size selection). On a small scale, it works quite well with recovery rates around 75-80% out of the box.

However, when I scale it up, I am reaching conditions where my recovery suddenly drops to below 60% while DNA:bead as well as elution buffer ratios remaining the same. Does the wash buffer volume play any role in the recovery? It seems as if I get better results with the lower volume batches than the high volume batches. I use 200uL 80% EtOH per wash. Would increasing the volume and/or amount of wash steps help in recovering more material?

 

thanks in advance

[Author]

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