rna from seeds

suter at VAX.MPIZ-KOELN.MPG.dbp.de suter at VAX.MPIZ-KOELN.MPG.dbp.de
Mon Nov 16 14:31:38 EST 1992

Subj:	rna from seeds

i am repeating this question on behalf of detlef becker, who is getting
rather frustrated....

he is trying to isolate mRNA from tobacco seeds. he has already tried two
classes of methods: guanidinium and phenol. basically what happens is that
everything turns an ugly kind of brown and is very sticky.
to circumvent this, he has added vitamin C and triton: total RNA looks 
better then.
but he is unable to isolate mRNA with either dynabeads or oligo dT cellulose...
both these matrices are attacked by his total RNA mix and partly
destroyed (this could be a pH effect: the Fe++ may dissolve from the
magnetobeads at low pH).

as i understood, he has learned from others in the mpi that they have 
practically the same problems with RNA from the seeds of other plants 
(rape i believe).

could anybody suggest what substances in seeds produce these problems ?
does anybody have a sure-fire method, that (s)he or a clever colleague
has used already ??? (yeah, yeah, he will accept a kit
also, as long as it works).

please post your answers to me or the newsgroup, i will forward them to detlef.



Clemens Suter-Crazzolara, PhD
Max-Planck-Institut fuer Zuechtungsforschung
Abteilung Genetische Grundlagen der Zuechtungsforschung
Carl-von-Linne Weg 10
5000 Koeln 30
Tel. xx49-221-5062.221           fax. xx49-221-5062.21
suter at vax.mpiz-koeln.mpg.dbp.de

More information about the Methods mailing list