I often use a speed vacuum concentrator to concentrate my DNA prior to restriction digests, DNA ligations, NGS library preparation or target captures.
Usually, my DNA is eluted in 10 mM Tris-Cl buffer without EDTA.
I am aware that speedvac'ing will inevitably increase impurities but most likely also salts. Would a higher concentration of Tris-Cl in the solution be a problem for the applications mentioned above, if we, say, concentrate the sample by a factor of 3-5? How do people usually deal with this?
Thanks for any ideas!
30 mM Tris is high enough concentration to actually buffer your DNA against any buffers you may add to it. If you are doing digests, your pH will most likely be off, unless your target pH is 8. You say you elute your DNA; if this is from glass bead or similar techniques where you elute by low salt, you can use water instead; no Tris needed to elute. Otherwise, you would need to dialyze or put through ion exchange, which I'm sure will dilute your sample too much. I don't have any other suggestions.
Hi,
SpeedVac of DNA will obviously increase the salt and Tris concentration which may interfere with your subsequent applications.
An alternative is to use autoclaved alkaline water, of pH around 8, to elute DNA. It works similarly to elution in Tris. The only downside is that very long term storage in water is not advised. However, if yo intend to use your eluted DNA in a couple of months, then using water is recommended.
Hope this helps!!!
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