difficult plant RNA prep

Kristin Bilyeu lastnamefirstinitial at missouri.edu
Thu Apr 12 16:59:14 EST 2001


Hello,

I have been attempting to isolate total RNA from whole Lotus japonicus
plants with no success and was wondering if anyone had suggestions.  My
latest efforts have been using the Pine Tree method of Chang, et al (Plant
Mol. Biol. Rep11:113-116. 1993).  My initial efforts with Trizol and
column-based purification schemes had problems with either polysaccharide
contamination or no recovery of RNA. The Chang method is fairly simple, yet
I am getting tiny pellets in the end.  I am going to go ahead and do an
RT-PCR with the whole pellet and try actin primers to see if anything was
really there (certainly not enough to check A260 or run on a gel).  Can
anyone give me some ideas?

Kristin Bilyeu
University of Missouri
Columbia, MO  65211





More information about the Methods mailing list