Dear Rosa
There are a number of protocols available, the TRIZOL method is a commonly used one, which we also have used a lot , some intro can be seen here http://openwetware.org/wiki/RNA_extraction_using_trizol/tri
Hello!
First of all I think you need to specify if you are planning on extracting RNA from fresh tissue or from fresh-frozen paraffin-embeded tissue! Once you specify that it will be easy for you to google the protocol you want - there is a large number of commercially available options!
Second thing to keep in mind regardless of where you want to isolate your RNA from is to make sure you do it in an RNAase free workflow!
Hope that helps!
normal breast tissue ? why take normal breast tissue. or autopsy ?
if fresh tissue or fresh frozen first tissue homogeizer system, extraction. Keep RNAase free water, and magnesium cloride to avoid effect of RNAase.
Dear all,
thanks a lot because of your answers,
I use fresh frozen tissue, RNase free materials and DEPC water,
I have seen protocols on google but they do not work for RNA extraction from normal breast tissue,
I used these protocol for extraction from TUMOR tissue and, that was great.
Dear Hasnaa, I used normal tissue against tumor tissue
for comparing expression profile
It is common in scientific article about cancer,
isn't it?
Regards
Dear Rosa
the most important thing to keep in mind is that breast tissue are densed with fatty tissues making it difficult to find really epithelial tissue
make sure to consult a pathologist in this case or go for laser micro-discection if it is paraffin embedded samples
good luck
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