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pombe - isolating tetrads

Michael Lichten lichten at helix.nih.gov
Fri Mar 15 12:37:05 EST 1996

In article <4ibtbl$8qg at masala.cc.uh.edu>, yoas at cfox.bchs.uh.edu (Karen
Yoas) wrote:

> Does anyone have advice or tips for using the micromanipulator for
> picking up
> asci?  (Besides lots and lots of patience...)

Make a new needle.  Make sure that your plates are dry (left unbagged >2
days after pouring).  Think only clean thoughts.

>  How do you prevent the
> cells from clumping when you put them on the plate?  I have tried
> vortexing and pipetting up and down, but I still have lots of cells
> clumped together.

One way to prevent cell clumping is to lightly sonicate.  We generally set
our sonicator on 0 (obviously not really 0) and sonicate for a few
seconds.  I have also found that the inclusion of 0.1% TWEEN-80 helps.

Michael Lichten
lichten at helix.nih.gov

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