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DNA extraction from FFPE tissue

DNA extraction FFPE PCR

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2 replies to this topic

#1 p.blaze53



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Posted 02 October 2015 - 04:08 AM

Hi everyone!


I've started DNA extraction from FFPE tissue and I've got low contcentration. I used QIAamp FFPE DNA Tissue Kit, tried it with 5 x 8um  &  5 x 6um sections and treat with xylen two times as well, but I couldn't get more DNA.


Does anybody have any idea, how I can increase the amount of DNA?


#2 2444256703



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Posted 04 November 2015 - 07:25 PM

any pictures? hope it is not so late

The secret to happiness is to keep setting yourself new challenges.

#3 merlav



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Posted 30 November 2015 - 12:25 PM

make sure that the paraffin is completely eliminated, how much time you incubate the tissue in protease? Make sure the tissue was completely homogenize (increase protease incubation time and/or use a homogenizer), warm the elution buffer, I can't remember what is the final volume of the elution buffer for this kit so what ever it is divide in half add to the column incubate for a minute (up to 5min), centrifuge, add the other half incubate again and centrifuge. Other tips for almost all qiagen extraction kits: never toss the column until you get a good concentration reading (sometimes if too much nucleic acids it get clogs if that is the problem add more elution buffer incubate 5 min (warm) and centrifuge (store columns at -20C until you are sure it can be toss), if the protocol says centrifuge 30 sec do it for a min, elute in 2 steps.  Always do the optional extra steps (those make a huge difference)

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