> | From: Virginia Berg <bergv at CHAOS.CNS.UNI.EDU>
> | To: plant-ed at net.bio.net> | Subject: plant physiology labs
> | Date: Monday, January 18, 1999 12:47 PM
> |
> | In an effort to persuade students (and faculty) that plant
physiology
has
> | something to offer students interested in biotechnology, I am trying
to
add
> | some experiments to the labs in my plant physiology course. Ideally
they
> | would be part of a single grand experiment to last up to 12 weeks,
mostly
> | spent waiting for the tissue to grow. Listed below are some of the
things I
> | would like to include. If we could do them with Fast Plants, we
could
get a
> | zillion seeds in a small space. Does anyone have labs already
invented,
or
> | suggestions for sources (human and text)?
> |
> | 1. making protoplasts (I have not done this much, and never in a
student
lab
> | setting)
> | 2. transforming cells (preferably with GFP, so detection is cheap)
> | 3. growing and selecting transformants
> | 4. tissue culture (maybe how to grow the transformed cells--we did
this
long
> | ago but stopped doing it as a lab)
> |
> | I recently reviewed a lab exercise in which seeds, rather than
tissue
> | chunks, were used for a tissue culture labs on the theory that it is
easier
> | to sterilize seeds and start with them, rather than sterilizing
tissue.
We
> | had a lot of contamination from the open air when we did tissue
culture
> | before, so I am planning to put together some boxes to keep the air
flow
> | down. If there are plans out there (cheap ones, please), I'd like
to
get
> them.
> |
I used the protoplasts isolation in a lab manual I will have to look
up. A
plant
biotechnology lab manual written by soemoneone in Minnesota. We used
cardboard
apple boxes laid on their sides, lined with aluminum foil and sprayed
just
before
use with 70% EtOH. EtOH sterilized all instruments before use, A/C'd
everything we
could. Got very good results. Of interest, two years later we had a
beautiful
tissue culture hood to use for the same set of experiments and got much
higher
contamination rates, go figure!!
Dave Starrett
