PCR of formalin fixed tissues

chelack at admin3 chelack at admin3
Thu Nov 24 03:41:34 EST 1994


I am also trying to set up PCR methods for formalin fixed tissue sections
and am concerned that the length of time the tissue was in formalin greatly affects the ability to amplify products.  In general I am deparaffinizing the
sections followed by freeze/thaw fracturing of the tissue and lengthy
proteinase K digestion then phenol chloroform extraction.  We are working
on the identification of M. bovis in elk samples.  The samples we have are
fixed for an unknown length of time and actually may vary in fixation time
among the samples.  So far I am able to obtain product from 4/7 samples
which leads me to believe that fixation time may be the culprit.  Does anyone
else out there have any other suggestions regarding this, or any other secret
tricks they wish to share?
                        Regards
                        BJC     Never worry about others trying to steal
                                your ideas, if they are any good at all
                                you will have to ram them down their throats.



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