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Isolation DNA and protein from a single sample


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#1 F.V

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Posted 28 April 2009 - 11:55 AM

Hello,

I need help from somebody that know how to extract DNA and protein from the same sample. I am working with animal tissue (lung).
If somebody has a protocol or a method to do that ( other than Trizol).
Thanks

FV

#2 bob1

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Posted 28 April 2009 - 04:42 PM

Your best bet is to divide your sample into portions and use one for DNA and the other for protein. Having said that, the first step in a DNA extraction is dissolving the tissue, so you could dissolve the tissue in a lysis buffer, take an aliquot for DNA extraction, add protease and go from there, and use the remainder for protein. Note: I haven't actually tried this, I usually just divide my samples.

#3 ntlfly

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Posted 28 April 2009 - 05:05 PM

Hi,
i have never done this myself, but i think you might try TRI reagent from Sigma,they apparently claim to use the same sample to retrieve protein,DNA and RNA.I have extracted RNA with Sigma, and it works like a dream.

Good luck

#4 F.V

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Posted 28 April 2009 - 06:13 PM

Your best bet is to divide your sample into portions and use one for DNA and the other for protein. Having said that, the first step in a DNA extraction is dissolving the tissue, so you could dissolve the tissue in a lysis buffer, take an aliquot for DNA extraction, add protease and go from there, and use the remainder for protein. Note: I haven't actually tried this, I usually just divide my samples.



Thanks Bob1.

The problem that I have is that I need to know the total amount of the DNA and Protein from the same tissue, I do not need for anything else the samples, just to know is one has more than the other.
What kind of lysis buffer do you use?
Thanks

#5 F.V

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Posted 28 April 2009 - 06:16 PM

Hi,
i have never done this myself, but i think you might try TRI reagent from Sigma,they apparently claim to use the same sample to retrieve protein,DNA and RNA.I have extracted RNA with Sigma, and it works like a dream.

Good luck


Hi ntlfly,
I think, it is the same that the trizol, the problem with that is when you need to work with the protein, it is very difficult to dissolve the pellet.

#6 bob1

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Posted 29 April 2009 - 04:26 PM

Thanks Bob1.

The problem that I have is that I need to know the total amount of the DNA and Protein from the same tissue, I do not need for anything else the samples, just to know is one has more than the other.
What kind of lysis buffer do you use?
Thanks

You could do this proportionally; cut the sample into bits and work out what fraction of the total weight each bit is, and then analyse from there.

For westerns, any of the usual lysis buffers will work, however as you are looking at total protein, you should probably do an industry standard test such as Kjeldahl or Dumas.




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