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paraffin from ffpe tissue samples refuse to dissolve in xylene


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#1 linmicro

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Posted 30 March 2011 - 03:39 PM

Hello
I'm trying to extract DNA from FFPE tissue shavings. I'm using xylene to disolve the paraffin but after several incubations and washes with it, the wax refuses to dissolve.
I was wondering if it is safe for the tissues to leave them in xylene overnight? and/or if there is a better protocol for it.
Thank you

#2 PostDocTrauma

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Posted 31 March 2011 - 07:33 AM

How big are the slices and how much xylene are you leaving in them?

we use 10ml of xylene for a 10umicron slice and leave it shaking overnight.

#3 linmicro

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Posted 31 March 2011 - 09:42 AM

How big are the slices and how much xylene are you leaving in them?

we use 10ml of xylene for a 10umicron slice and leave it shaking overnight.

I have samples weighting between 50 and 60 mg (tissue plus wax) in eppendorf tubes. These are pieces that were shaved from a block of ffpe tissue.
I'm using 2mls of xylene to dissolve the paraffin, because a protocol that I'm following calls for 1ml of xylene to be used in 25 mg of ffpe tissue sample.
But I might need to use more xylene according to what you mentioned.
Thank you so much for your answer and help




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