Green Gene Project

dkim at unm.edu dkim at unm.edu
Mon May 23 13:10:53 EST 1994


Richard Dana asks:
[quote]
I want methods for isolating RNA from plant leaves, stems, roots and the
best storage conditions until the RNA can be studied. Is plant RNA any
different than other species and is there anything unique about
stabilizing isolated plant RNA.
[end quote]

I don't think there should be any difference in the RNA as it is. I am told,
however, that plants have a variety of nucleases which are active under
conditions of extreme cold or heat, unlike mammalian cells which maintain
tighter control over their environment (in the whole organism).

I isolate RNA from suspension cultures, but the idea should be the same for
tissues:
I use a filter paper to dry the cells a bit, then quick-freeze in liquid
nitrogen. I then grind to a fine powder in a mortar and pestle, then add
"Solution D" from the Acid guanidinium RNA isolation procedure of 
Chomszinski and Sacchi (I never spell it right :)) to the cell powder. After
clearing cell debris, the RNA isolation is right out of the published procedure.

Cleared lysates can be stored at 4 C for years, I am told. I have stored the
redissolved RNA (in DEPC-water) at -20 C for weeks with no apparent ill 
effects.

Daniel Kim



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