[Histonet] RNA isolation form stained slides

Vanessa Orsini vanessaorsini <@t> msn.com
Fri Nov 2 04:42:46 CDT 2012







Hello,

I need to extract RNA for a RT-PCR after Laser Micro
Dissection on xGal stained slides.

I tried using sections from unfixed frozen organs. I fixed the
sections in EtOH70% for 10min and then I stained them with xGal for 3h at 37°C.
After air drying I cut out with the LCM and extract RNA with the PicoPure kit
from Applied Biosystem. So far I didn’t manage to get enough RNA. 

I tried to add RNase inhibitors to all the solutions but it
didn’t help.

 

Any idea/suggestion?

Do someone think it would be better to do a LacZ antibody staining
on FFPE sections and extract RNA with an appropriate kit? The RNase would they
be less active?

 

Thank in advance for any help you can give me J Vanessa

 		 	   		  


More information about the Histonet mailing list