I have a liver biopsy which I want to screen for the expression of liver carcinoma related proteins. It is 10 mg of tissue embedded in paraffin for making microscopic slides. I want to extract mRNA for expression analysis by pyro-sequencing. The sample is unique, so I want to be sure that the procedure gives results (data will be linked to MRI, CT scans and extended blood chemistry, which are already available).
I have no direct experience but I know there are different kits which seem to give good results. I suggest you to check in the literature some specific information.
Hi, i can suggest you ReliaPrep™ FFPE Total RNA Miniprep System, this kit could be an option to extract RNA from your samples. Here below the link to kit specs:
http://www.promega.com/products/dna-and-rna-purification/rna-purification/reliaprep-ffpe-total-rna-miniprep-system/
Hope to be of help.
We have used very recently a kit available from Roche on sections from liver specimens embedded in paraffin (Roche code 04823125001, High Pure FFPE microkit). The kit was indicated as the best one available by a number of colleguaes. We were not so much happy of results (very low levels if any and not clean mRNA) but we honestly (because of very small amount of human materials available) tried to obtain mRNA from 2-3 sections only (3-4 um thick), that is presumably much less than 10 mg you have. The kit is easy to use and even unexspensive but you may need to proof it first on less "precious" samples.
Hi
We use Qiagen's RNEASY FFPE kit for this sort of work. We'd receommend (since you're working from a biopsy) 3-4 sections of 5-6um thickness but of course your downstream needs might mean you need more. Catalogue number is 74404.
Thanks for the replies. I have some useful suggestions which I will follow and test. Keep you updated.
Thanks! Harrie
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