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Old 01-31-2017, 03:00 PM   #1
herter
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Default Magnesium + heat = Shear DNA?

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!
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Old 01-31-2017, 06:22 PM   #2
ECO
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Without a 2' -OH, the DNA won't shear.

Magnesium is in high concentrations in every PCR buffer also.
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Old 01-31-2017, 08:02 PM   #3
nucacidhunter
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If you are looking for metal ion based DNA shearing see the following:

https://beng.soe.ucsc.edu/sites/defa...esis_Final.pdf
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Old 02-01-2017, 09:41 AM   #4
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Anyone tried this? ^^

Seems gloriously too good to be true.
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Old 02-01-2017, 10:01 AM   #5
herter
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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.
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Old 02-01-2017, 11:53 AM   #6
nucacidhunter
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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.
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