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
-
CRISPR/Cas9 sparked the gene editing revolution for both research and therapeutics.1 But this system still showed severe issues that limited its applications. The most prominent were the heavy reliance on PAM sequences, delivery limitations, double-stranded breaks that prompt unintended edits and cell death, and editing inefficiency (both in targeting and in knock-in reliability).
Despite this, “CRISPR helped turn genome editing from a specialized technique into...07-31-2026, 11:01 AM -
Proteomics platforms are evolving rapidly, with advances in mass spectrometry and affinity-based approaches expanding what researchers can detect and at what scale. As the field moves toward deeper proteome coverage and clinical applications, scientists face an increasingly complex landscape of tools. This article will explore how researchers are navigating these choices to find the right platform for their work.
The systematic characterization of the human proteome has...07-20-2026, 11:48 AM -
Genomics studies in neuroscience face a special challenge due to the brain’s complexity and scarcity of samples. Mapping changes in cell type and state using conventional next-generation sequencing methods remains challenging. Advances in technologies like single-cell sequencing, spatial transcriptomics, and long-read sequencing have opened the door to deeper studies of the brain and diseases like Alzheimer’s, amyotrophic lateral sclerosis (ALS), and schizophrenia.
...07-09-2026, 11:10 AM
Comment