What do you think?

Koen De Smet kds30
Mon Jan 8 09:37:22 EST 1996


syamada at sb.gunma-u.ac.jp wrote:
>I'm now extracting m-RNA from some digestive tract tissues. I cannnot get
>good results .I think it may be the influence of the tissue RNAse.How can
>I efficientry reduce the RNAse activity?  I use ''trizole'' ,one of the
>phenol extraction  kit,now. Specimen is cut out and immediatry frozen
>with liquid N2,then mixed with ''trizole'' by homogenizer.
> I think it may be better to prepare the  specimen  freeze dryed and then
>infiltrate some fluid ,such as ethanol,before  using RNA extraction kit.
>What do you think about this method. Drying could aggregate the RNA ,and
>disturb collecting it ?
> If some one know about this.Please tell me.

I have used the method from P. Chomczynski & N. Sacchi, Anal. Biochem., 162, 156-159 (1987) succesfully for extracting mRNA from several tissues, including pancreas which is full of RNases. It uses guanidinium thiocycanate followed by phenol extraction. I never froze the tissues. I collected them, washed in saline and then put them in the guanidinium thiocycanate solution. I chopped it in little bits and then homogenised them.
The RNA was used succesfully for Northerns and for a cDNA library.

Koen De Smet





More information about the Methods mailing list