Especially when it comes to Illumina libraries, why is it commonly recommended to Speed-Vac without heat? Is this to prevent (or slow down) potential DNase activity?
-
-
I believe this recommendation usually applies to RNA samples - which are more heat sensitive.
Depending on the buffer (some labs might use be pure H2O) and the lengths of fragments, using low temperatures (e.g. room temperature) might avoid denaturation of DNA samples. -
Thanks for the response, luc! I'm currently working through a ddRAD protocol (my second library). Long story short I have digested DNA w/ ligated adapters. I bead-cleaned them after pooling, but eluted with a large volume of HPLC H2O and Speed-Vacced to reduce the volume (from ~ 150 uL to ~ 30 uL).
Our normal vacuum pump is broken so I'm using a weaker replacement. It was taking forever so I upped the temp to ~ 45 C.
...and then, as often happens, I made myself paranoid. But I'm also genuinely curious about this recommendation. So thanks!Comment
-
High temperature can also impact AT rich sequences and degrade them.Comment
-
Interesting. I'll look into this a bit more, thanks.Originally posted by huguesparri View PostComment
-
Very, very likely your libraries will be perfectly fine. 45C should not be a problem, even in pure H2O. This is my guess, though.Comment
-
Tumors contain complex mixtures of cell populations that evolve in response to treatment and host-specific factors. Tissue biopsies provide only static snapshots of a tumor’s state. But what if we could follow that evolution over time using a minimally invasive technique?
Liquid biopsies can detect tumor-derived material circulating in the bloodstream, providing a way to study treatment response, detect emerging resistance, and discover new biomarkers. Circulating...10-14-2026, 09:55 AM -
Researchers using sequencing and genomics tools often have to make trade-offs. They can choose between speed or scale, short reads or long-range information, or targeted panels or a view of the whole transcriptome. New technologies that have been released this year are built to address those tough choices.
We asked six companies the same four questions to learn about their latest products. The new technologies bring a lot to the table, including rethinking sequencing...
Comment