Help - mRNA from frozen starfish

Marianna Max drmax at casbah.acns.nwu.edu
Fri Apr 28 06:12:10 EST 1995


In article <3npqs8$4nf at server.st.usm.edu>,
Shiao Y. Wang <sywang at whale.st.usm.edu> wrote:
>Richard Waring (waring at astro.ocis.temple.edu) wrote:
>: We want to purify mRNA from frozen (-70) starfish ovaries. We have  
>: experience purifying mRNA from lyophilized fungi using various methods.
>: We plan to use a guanidinium isothiocyanate method but would appreciate 
>: suggestions for getting from frozen chunks of tissue to the point where we can
>: homogenise in extn buffer.  Spawning of the starfish is seasonal and so 
>: we only have limited amounts of material to experiment with.
>
>I would suggest pulverizing the frozen ovaries in liquid nitrogen first.
>Add the pulverized ovaries to G-SCN soln directly and homogenize quickly.
>
>Shiao Wang
>University of Southern Mississippi
>

I wouldn't bother pulverizing. Make frozen chunks small enough to fit into
whatever tube you want to spin in and put them directly into G-SCN solution on
ice and polytron (still on ice). We do this with frozen retinal or brain
tissue all the time.

Max





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