I have brain frozen mouse brain areas (frontal cortex, hippocampus and hypothalamus) and I would like to extract RNA from them using the Trizol or the RNeasy kit. Is it sufficient to use a insulin syringe and needle to homogenize these brain pieces (mm for less than 20 mg tissue) or is it better to use a tissue lyser? What approach will lead to RNA samples with the highest RIN?
Thanks
Hi Ilario,
Trizol is a very good method for RNA purification that provides very good RIN values. I have always used either a tissue homogenizer (polytron) or a mortar and pestle. Both methods give good results. However if money is not an issue I would recomend you to use try one of the qiagen RNA purification kits that also provide very good results.
Hope this helps,
Ezequiel
Syringe and needle is sufficient, and the advantage is that you are only using disposable material, avoiding possible cross-contaminations. Be careful with frozen tissues: RNase will start acting as soon as they melt. So I prefer to disrupt fresh tissues, and to freeze the homogenized lysate (high salt buffer inhibits RNase) if I cannot do the extraction the same day.
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