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Seed selection after transformation

matej lexa m-lexa at students.uiuc.edu
Tue Dec 8 18:13:55 EST 1998


Keiper <Keipern at esvax.es.dupont.com> writes:

>I'm looking for protocols for the sterilization of A.t. seeds harvested
>after a vacuum infiltration with agro.  I have been sterilizing with 50%
>bleach/0.02%trition x-100 for 7 mins, but have been getting a lot of
>contamination on my plates.  Any suggestions?
>Thanks,
>Natalie Keiper-Hrynko

I have been using the following modification quite successfully:
After the above treatment (or a similar treatment using 70% alcohol/triton)
drain the tubes with seeds of most of the liquid, squirt in 95% alcohol
and immediately dump the contents on a clean piece of filter paper.
After alcohol evaporates, transfer seeds from the filter onto agar.
The whole procedure is done in a sterile environment - like a laminar flow
hood.


--
Matej Lexa                  e-mail: m-lexa at uiuc.edu
Dept. Plant Biology         home  : http://www.students.uiuc.edu/~m-lexa/
University of Illinois      fun   : http://net-cities.com/
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