DNA sample contaminated with lipids - (Aug/30/2010 )

Hello All
i have used salting out method for extraction of Genomic DNA from urinary bladder of mice but as the tissue contain large amounts of fats.
so the Extracted DNA is contaminated with lipid.

HOW could i get rid of those fats from my DNA sample???

Lipids are very soluble in chloroform, so a simple chloroform extraction will remove them.

phage434 on Tue Aug 31 02:11:02 2010 said:

I agree. That would solve the problem

phage434 on Tue Aug 31 02:11:02 2010 said:

DNA now in TE buffer how could i apply chloroform extraction on it???

The same as you would for any other chloroform extraction - add 1 volume of chloroform to the sample, shake or vortex vigourously for 30 s, then centrifuge to separate layers...

bob1 on Thu Sep 2 23:56:46 2010 said:

without need for phenol:chloroform step at first?

jehane on Fri Sep 3 00:04:28 2010 said:

Yes, just add chloroform.

The purpose of phenol is to denature proteins. Since proteins aren't a problem here, there is no need for phenol:chloroform mixture.

However if you don't have chloroform and only have phenol-chloroform, you can use phenol-chloroform mixture instead.

I'd recommend an ethanol precipitation and resuspension in TE following the chloroform extraction.

Also, there are kits for extraction of DNA or RNA from high-lipid tissues. I use the RNeasy Lipid Tissue Mini Kit for RNA, but there should be some for DNA as well.