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Old 11-04-2016, 02:39 AM   #16
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Location: Germany

Join Date: Aug 2012
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Originally Posted by chadn737 View Post
I have used Turbo DNase followed by cleanup on Qiagen columns. The Turbo DNase kit does provide an inactivation agent that is supposed to remove the enzyme, but time and again I find that the inactivation agent fails to adequately remove the enzyme (I have been using it for ~ 5 years now). For more routine uses, I'll use the inactivation agent, but for my more valuable samples, I always follow up with cleanup on a column.
This thread is rather old, so I'm not sure if I'll be able to get a response, but here goes...

When you do the column purification to remove the DNase enzyme, do you add buffers (which buffer/what volume?) to your RNA sample before passing it though the column and proceed with the same protocol you used initially to extract the RNA? (With Qiagen Mini kit: RLT, RW1, RPE.) I need to remove some DNA from my RNA samples and I'm a bit concerned that you said the inactivation agent isn't completely removed. I want to make sure the cDNA created from my RNA libraries isn't automatically digested. Thanks for your input.
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