pombe - isolating tetrads

Singer Instruments yeast at singerinst.co.uk
Tue Mar 26 09:10:34 EST 1996


In article <lichten-1503961237050001 at 128.231.218.92>, Michael Lichten
<lichten at helix.nih.gov> writes
>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.
>
Reduce the concentration of your inoculum and be careful how much liquid
you deposit when you streak.

We make and sell needles for ascus dissection; they work well with pombe
tetrads which are a little "sticky". Email me for details.

Carl Singer
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