micromanipulation of veg. cells

Kevin A. Morano kmorano at umich.edu
Mon Aug 5 11:07:12 EST 1996


Hi all,

I'm trying to manipulate vegetative cells using a sick Rainin apparatus
and am having trouble separating daughter and mother cells.  I am doing
all of this on an inverted YPD plate and can move the cells around no
problem.  The hangup occurs when I try to separate a large bud from mom or
worse, when the bud fails to pinch off before a new bud emerges.  For my
analysis I really need to be able to recognize who budded from whom and
get them away ASAP.  Does anyone have advice on encouraging daughter yeast
to move out?  WHile I'm at it, does anyone know why they stay clumped for
so long and continuously make new buds before separating?

Thanks in advance for any advice!

Kevin



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