[Histonet] LCM & archival FFPE tissue

Margaryan, Naira NMargaryan <@t> childrensmemorial.org
Tue Feb 22 13:26:47 CST 2011

Hi all,

I have a couple questions I would like to throw out to the experts:

1)      1) Is it possible to extract RNA and DNA for gene expression purpose after usual IHC procedure on 4um sections of archival FFPE breast tumor tissue (If tissue was in citrate buffer, pH 6.0; several blocking steps; one hour in primary Ab (less then one hour will not work); with immunoperoxidase-DAB detection follow the Hematoxylin?
2) Or I need only H&E?
2)      3) Is 4-5 um section good enough?
3)      4) How many sections need to have really good and highest yields?
4)      5) What temperature of Proteinase K is need 42º or 60ºC?
I've the LCM system from Arcturus and the Pico Pure Kit.

Any suggestions?

Thanks in advance,

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