plant rna extraction : purity problems

[P_Kianian at ndsu.nodak.edu]

Hi

We also have noticed the same problem when isolating 
RNA from roots of various wheat and durum cultivars 
using the LiCl method.  The only samples we had success 
with were young roots from germinating seeds.  We 
currently don't have an answer to this problem but 
thought the root-tips of the "older" plant might give 
us more success in our RNA isolation. 

Penny Kianian 



Quoting Sam Lievens <salie at gengenp.rug.ac.be>:

> hi all,
> 
> i'm having serious problems getting pure rna out of 
the roots of our
> plant. it's a tropical legume. getting rna from the 
aerial parts is no
> problem (od 260/280 of 2.0 and 260/230 of about 2.3), 
pure enough to
> prepare cdna of.
> but using the same protocol (licl based method) on 
roots gives me dirty
> rna (od 260/280 still around 1.8 but 260/230 of only 
1-1.5 so i guess
> caused by polysaccharide contamination). i tried 
other methods (trizol,
> quiagen rneasy columns) but it doesn't help.
> so two questions: how can i get pure rna and how can 
i further purify
> the dirty rna i already have (i already precipitated 
it again with licl,
> doesn't help; i did 2 fenol/chloroform extractions 
and ethanol
> precipitation, doesn't help neither).
> 
> sam
> 
> --
> 
========================================================
===
> Sam Lievens
> 
> DEPARTMENT OF GENETICS       -       UNIVERSITY OF 
GENT/VIB
> K.L. Ledeganckstraat 35    -    9000 Gent    -      
Belgium
> tel: +32(0)92645183                     fax: +32(0)
92645349
> salie at gengenp.rug.ac.be  
http://www.plantgenetics.rug.ac.be
> 
========================================================
===
> 
> 
> 
> 


---