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DNA isolation from nuclei - (Jul/20/2013 )

Hello! Has anyone ever isolated mammalian DNA from nuclei? I can make nuclei really easily for DNase Hypersensitivity experiments and I receive significant yields at the end, so I thought it should be straightforward to isolate DNA directly from nuclei rather than from a Qiagen kit. Thanks!

-hotstuffdb22-

not sure what you mean? Nobody forces you to use a qiagen kit... So I dont see what your problem is or what you question is? Do you mean whether you could use another type of DNA extraction system than the qiagen kit?

-pito-

Thanks for the reply.
Normally, I isolate DNA from 5 million cells or digest tissue overnight, either with a qiagen kit or routine phenol-chloroform/ethanol-ppt. Just curious about whether DNA from nuclei can be isolated using phenol-based method.

-hotstuffdb22-

hotstuffdb22 on Sat Jul 20 16:41:37 2013 said:


Thanks for the reply.
Normally, I isolate DNA from 5 million cells or digest tissue overnight, either with a qiagen kit or routine phenol-chloroform/ethanol-ppt. Just curious about whether DNA from nuclei can be isolated using phenol-based method.

I am a bit confused. I dont really understand your questions.... You do a DNA extraction with the phenol-based method, so of course you can ge the DNA from the nuclei (where else would the DNA come from?) I am not sure I understand you.... You work with tissue, so I assume its from a eukaryotic organism?

-pito-

I apologize for the confusion. I'm not being clear here. Thanks for the feedback!

-hotstuffdb22-

hotstuffdb22 on Sun Jul 21 20:48:25 2013 said:


I apologize for the confusion. I'm not being clear here. Thanks for the feedback!
Just to be sure: the DNA is in the nuclei (exceptions not mentioned) , so yes, you already did get DNA from the nucleus... ANd phenol(-chloroform) extraction is good.. its even better than the kits if you work with low amounts of DNA (better to work with isopropanol than for the cleaning step).

-pito-