RNA degrade - RNA degrade during in DNase treatment (Aug/02/2005 )

Hi, guys;
I meet a problem. When I try to remove the DNA from my RNA sample(looks good from gel-running) with DNase, the RNA was degraded(Please look the image!)? Who can tell what reasons can cause it?
Thanks a lot!
Clint

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If it looks fine before your DNase reaction, then your DNase may be contaminated with RNases. Also, are you performing your DNase reaction immediately after you look at your RNA on a gel? If not, then it may be your RNA storage method that is causing the degradation (we all know how finicky RNA can be). If you are indeed performing the DNase reaction immediately after you check out your RNA on a gel (and it looks fine), then try a different batch of DNase and see if that helps.

Best of luck,
Hank

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This recently happened to me as well - and I'm pretty sure that the DNase was contaminated. That may also be what happened in your case.

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Thanks, guys.
I am doing another batch of RNA isolation. I will try another DNase and see what will happen!

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Make sure that you do a parallel experiment where you incubate your RNA under the same conditions (buffer, temp, etc) as the DNase digestion except don't any DNase. The RNase contamination may be coming from another source other than the DNase.

Daniel

DNA sequencing analysis software

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