non-isotopic in situ PCR: biotin or dig?

Gregory E. Gonye PhD gonyege at lldmpc.dnet.dupont.com
Wed Apr 12 16:35:39 EST 1995


Hello fellow surfers,

We are planning a series of experiments which will require combining in situ
PCR and immunohistochemistry on the same thin section.  We would like to
stick with HRP or AP driven reactions if possible (directly conjugated
secondary antibodies) or double-labelled immunofluorescence (one for
beta-gal and one for the PCR product), sssssoooooo:  How would you do it? 
end label or incorperate? Digoxygenin (sp?) or biotin?  I'm guessing
Amplitaq will like one better then the other and that incorperation will be
more sensitive than end-labelled.  But I've never done this before and have
not been able to find this info in the normal places.  Help please.  If it's
not too much trouble, please mail directly to me (and post as well as this
seems like very useful discussion).  Thanks in advance {:-)   G.



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