Protocol for RNA preparation from Formalin Fixed tissues - (Dec/24/2008 )
Dear Friends,
I am eager to extract total RNA from Formalin fixed tissues (either paraffin embedded or not).
Unfortunately, I can not get a commercial kit so I would like to prepare all the reagents by my self.
I have basic reagents for RNA preparation, such as Trizol reagent etc, so I can use them in the procedure.
Could anybody let me know a reference or give me a protocol for it without using commercial kit?
I am dealing with mammalian tissues.
Thank you very much for your help in advance.
TK
I am eager to extract total RNA from Formalin fixed tissues (either paraffin embedded or not).
Unfortunately, I can not get a commercial kit so I would like to prepare all the reagents by my self.
I have basic reagents for RNA preparation, such as Trizol reagent etc, so I can use them in the procedure.
Could anybody let me know a reference or give me a protocol for it without using commercial kit?
I am dealing with mammalian tissues.
Thank you very much for your help in advance.
TK
Hi,
I know an article of extraction of viralRNA from Formalin-fixed, paraffin-embedded tissue blocks, so i think its a free article, if it don´t please tell me. I don´t know if it´s possible to send the pdf from here or by e-mail. Any way the article it´s called:
Development of a semi-nested reverse transcription polymerase
chain reaction assay for the retrospective diagnosis of canine
distemper virus infection
James B. Stanton, Steven Poet, Salvatore Frasca, Jr., Dorothee Bienzle, Corrie C. Brown
J Vet Diagn Invest 14:47–52 (2002)
I hope this help you, and check the References
Is it that you are not allowed to get a kit--or you can't find a kit? I do know of a great one for these types of samples that just came out this year. Nucleospin RNA XS. It is specifically designed for these types of samples and allow a 5ul elution. I have used it on some tiny marine samples. I believe it can purify as little as 1-10 cells . It is a simple spin kit protocol. Good luck.
I am eager to extract total RNA from Formalin fixed tissues (either paraffin embedded or not).
Unfortunately, I can not get a commercial kit so I would like to prepare all the reagents by my self.
I have basic reagents for RNA preparation, such as Trizol reagent etc, so I can use them in the procedure.
Could anybody let me know a reference or give me a protocol for it without using commercial kit?
I am dealing with mammalian tissues.
Thank you very much for your help in advance.
TK
Thank you very much to BiolMolMex and Lunchbox.
Unfortunately, we have limitation of budget to buy a kit for this experiment at this moment, especially this is kind of trial experiment for us now. But, thanks for the information of the kit. I will keep in mind till the time I can buy a kit.
Thank you for the information of the reference. I could download it and checked briefly.
I wonder some point on this protocol whether they extracted nucleic acid using acidic phenol or not (no mention in the materials and methods). No description of DNase treatment, either. Maybe, I will use Trizol reagent to extract RNA and use primers which amplify the sequence containing intron.
The information for ProK treatment is very helpful for me.
Thanks again!