Please, help with RNA extraction.

[Thibaud Le Mouel]

>I have problems when I try to isolate total RNA from mouse liver or brain by
>Chomczynsky method, because the two phases I should obtain after adding
>phenol and chloroform doesn't separate during centrifugation. This
>difficults the extraction and makes the efficiency to be very low.Does
>anybody know why this happens and how to avoid? Is this method good enough
>to extract RNA from those tissues?
>
>I would appreciate your comments a lot.
>Thank you,
>
>Maria
>
>melia at ar.vhebron.es

Hi,

I think there is too much lipids in the liver and perhaps you put too much
liver for your extraction. We use only 1 mg of liver for 5 ml of extraction
buffer and  we make 2 phenol/chloroform because of lipids

Hope that can help you.

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