RNA extraction from FFPE tissue
Posted by: JosephineHH
Date: January 26, 2012 12:15AM
Is there anyone in this forum, which has experience with RNA extraction from formalin-fixed, paraffin-embedded tissues? I use "Qiagens RNeasy FFPE kit"with 2 slices of FFPE tissue per reaction, each with a size of 10um.
Following RNA extration I use the RNA as template for reverse transcription with "Qiagen QuantiTect RT kit", with 100ng RNA as starting template in a total volume of 20ul.
Of the 20ul cDNA I use 3 ul for further processing in quantitative PCR using "Qiagen, Rotor-Gene SYBR Green kit".
The RNA yield is very low and I dont know whether is it due to low starting material, too short deparaffination period with xylen or if I need a higher cDNA concentration in my qPCR.
I will proceed my experiments and try to regulate these thoughts, but in the meantime I am curious to know, whether anyone of you have any experience with RNA isolation from FFPE tissue, and if you have any experience with Qiagens products?
Thanks in advance!