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I know in some RNA-Seq protocols magnesium and heat are used to shear the RNA. I'm curious if magnesium and heat will also shear DNA?
Thanks!
Thanks!
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Without a 2' -OH, the DNA won't shear.
Magnesium is in high concentrations in every PCR buffer also. -
If you are looking for metal ion based DNA shearing see the following:
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Anyone tried this? ^^
Seems gloriously too good to be true.Comment
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Thanks for the replies! Any recommendations on chemical or enzymatic shearing for DNA and RNA (TNA) at the same time? We do mechanical shearing now for TNA, but hoping to find a faster/easier approach.Comment
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It is possible to use combination of DNA fragmenting enzyme such as NEB fragmentase with RNase III. The issue would be buffer and incubation temperature and time requirement which can be overcome by treating with one enzyme followed by dilution and adding other enzyme and buffer without a need for clean up in between and optimising enzyme quantity.Comment
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DNA Fragmentation at Taq Premix
recently I have tried it
Add 400ng Lambda DNA(48.5K) into 1X Taq Premix with 2mM MgCl2,
it shows clearly fragmentation more than 1min at 95C if more 4min median of fragment is around several Kilo; 95C 16min most of fragment is small than 1K.Comment
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I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.
Here are nine questions we think about, in roughly the order they matter, before...06-18-2026, 07:11 AM -
Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.
The first step is collection, followed by preservation and sample preparation for analysis. Most scientists overlook those steps, but not being careful might just be skewing the experiment’s results.
...06-02-2026, 10:05 AM
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