Getting seeds from transgen
comai at U.WASHINGTON.EDU
Thu Sep 5 19:30:39 EST 1991
re: from dish to soil, another recipe...
We do our transformation work with ecotype No-0. We usually obtain some
pods from Kan-R shoots growing on AGM plates. Here is the recipe:
0.5X Murashige and Skoog salt mix (Gibco 1117)
0.6% Gibco Phytagar
1X Gamborg B5 vitamins (add after autoclaving)
pour in 10x2.5 cm plates
These plates are sealed with porous surgical tape (3M 1530-1). While
most shoots seem to produce seed, we prefer to transfer them to soil
where the seed yield is much higher and seeds are easier to collect.
According to our experience, failure to establish a rooted shoot in soil
is due to damage or loss of root system during removal of the plant from
the agar medium. The following protocol solves this problem and gives
over 90% survival of transplants. Applying care, one can actually get
100% survival. By the way, this method also works for seedlings selected
by whatever screen on petri dishes.
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