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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