[Histonet] RNA and DNA yields from Laser Microdissection

[nefff <@t> staff.uni-marburg.de]

Hi Carol!
My experience is, the yield depends on the protocol you use. I've done  
the LCM system from arcturus and the PALM-System from Zeiss. The  
highest yields had been  for RNA and DNA by using the Pico Pure Kit  
(MSD) while doing LCM. This had been frozen sections. We isolated DNA  
from FFPE-Tissue from PALM and LCM by a conventional Proteinase K  
digest and phenol extraction.
There is a paper of Fend and Specht at the Am. J. of Pathology, 2002  
or 2003 describing these protocols.
As long as the histopathologist or the tech are able to identify the  
"cell/tissue" of interest, it doesn't influence the yield/quality of  
the RNA/DNA.

Frauke


Zitat von "Barone, Carol " <cbarone <@t> NEMOURS.ORG>:

> Histonet: I have a couple questions I would like to throw out to the experts:
>
> 1. Do you think higher yields of DNA /RNA in LMD are related to   
> protocol, method differences from one instrument to another ( i.e.   
> catapult verses,, gravity drop, verses,
> arcturus melt system...or more related to histologist v. research   
> tech handling of the sample?
>
> 2. How do you feel about frozen verses paraffin as related to yield   
> and quality in LMD/LCM?
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