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-   -   DNA extraction from cells fixed in 4% PFA (http://www.molecularstation.com/forum/dna-techniques/87792-dna-extraction-cells-fixed-4%25-pfa.html)

Mastah 04-15-2013 06:26 AM

DNA extraction from cells fixed in 4% PFA
 
Hey! I have a problem... Does anyone have a good protocol to extract DNA from cells (like cancers cells or stem cells) fixed with PFA 4% (Paraformaldehyde) ??? It's only about 5000-15000 cells. My protocol... 1h in 55oC - 100ml Tis, 20ml proteinsae K and 200ml LT is not good :frusty:

admin 04-24-2013 06:45 PM

Re: DNA extraction from cells fixed in 4% PFA
 
What are you getting with your protocol in terms of DNA quality and quantity?

Mastah 05-06-2013 11:32 AM

Re: DNA extraction from cells fixed in 4% PFA
 
With my protocol... there's nothing. I get a value about 1-2 ng/ul (quality: 0; 0,5).
I increased the amount of proteinase K and length of incubation ... still nothing.


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