DNA extraction from FFPE tissue - (Oct/02/2015 )
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?
any pictures? hope it is not so late
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)