rna from seeds, some more problems

suter at de.dbp.MPG.MPIZ-KOELN.VAX suter at de.dbp.MPG.MPIZ-KOELN.VAX
Mon Aug 31 07:36:09 EST 1992


 brian tague wrote:

<Subj:	Isolation of RNa from seeds
<
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<==================
<Does anyone have a reliable method for the isolation of RNA from dry seeds
<and/or yellow siliques. With the standard RNA prep (SDS/LiCl/phenol/chloroform)
<some kind of contaminant comes through with the RNA. Procedures for the removal
<of polysaccharides (1M NaCl ppt @ 0C, acid ppt) don't remove it. Any hints or
<suggestions would be greatly appreciated. Thanks in advance.
<-- Brian W. Tague

we are currently having the same problem with RNA isolated from tobacco seeds:
mRNA is prepared according to your method and purified further with dyna-
beads. Resulting RNA is rather brown (in contrast to leaf rna) and cannot
be used to efficiently produce a cDNA library (again in contrast to leaf rna).

is there some oxidating agent in seeds ? we notice that seeds, freshly opened 
are white, but turn brownish within 30 minutes.

any hints welcome.

clemens
----------------------------------> address <----------------------------------
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
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