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DNA isolation from nuclei

DNA Nuclei method isolation phenol-chloroform

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5 replies to this topic

#1 hotstuffdb22

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Posted 20 July 2013 - 07:28 AM

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!

#2 pito

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Posted 20 July 2013 - 08:21 AM

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?

If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#3 hotstuffdb22

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Posted 20 July 2013 - 08:41 AM

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.

#4 pito

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Posted 20 July 2013 - 09:25 AM

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?

If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#5 hotstuffdb22

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Posted 21 July 2013 - 12:48 PM

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

#6 pito

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Posted 21 July 2013 - 01:03 PM

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).

If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.






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