arabidopsis seeds

Kathy Wilhelm wilhelmk at studentw.msu.edu
Fri May 20 20:15:00 EST 1994


In Article <2rirqs$rjo at mserv1.dl.ac.uk> "STEPHEN A ROLFE <S.Rolfe at sheffield.ac.uk>" says:
> Dear netters
> 
> Please excuse this extremely lo-tech question.  What is the best way
> to evenly disperse seeds on an agar plate for kanamycin selection?  I
> sterilise the seed in 10% bleach with a drop of detergent followed by
> several water washes.  This works fine - I don't get bugs on my
> plates.  However the seeds do aggregate when placed on the plate
> which is a bit of a pain.  I wondered about suspending the seeds
> in a .1% agar solution but this is a bit inconvenient.  If anyone has
> a wonderful idea then I would be very grateful to hear it.  I am
> slightly wary of using dry seed sterilised by microwaves after
> previous debates on this matter.
> 
> Thank you in advance
> 
> Steve Rolfe
> 
> s.rolfe at sheffield.ac.uk
I've tried several methods of spreading seeds, including using 0.1%
 agarose, pouring them onto the plate in water-based "top agar" &
 spreading them around with a bent glass rod the way we do bacteria. 
 I haven't been satisfied with the evenness of the spread for any of these.
My tried & true method is to take up the seeds in the residual water left
 from the sterilization and, keeping a gentle, steady pressue on the pipetman,
let the seeds out one by one.  It's tedious, but its the only method that's really 
given me a good spread.
Best wishes!
                                Kathy_|_
                                      |
> 



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