I am preparing Nuclear Matrix, the insoluble filamentous fraction of the nucleus. Here are the steps:
Isolate nuclei
Digest DNA with dnase and detergent
Extract in 400mM and 2M NaCl respectively
dnase should come out in the supernatant after digestion or high salt wash. But it is stuck in the nuclear matrix which could alter my results. What is the problem?
Dmitry Guschin
Hi Ashish, what are you going to investigate? Could you simply heat-inactivate DNAse1? If you next step is some sort of PCR, heat inactivation should not be a problem.
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Ashish Bihani
Hi. Thanks for response.
No. I want to look at the ultrastructure of the residual complex. The activity is not a problem. DNAse could cause formation of faux structures.
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