Yeast clone recovery from glycerol stock

Anna s990192 at mailserv.cuhk.edu.hk
Mon Sep 11 13:26:52 EST 2000

Hi All,
Just wonder if there is a good method to recover the yeast clones from a
glycerol stock?
The glycerol stocks were prepared 5 months ago, they are clones screened
by Yeast Two-Hybrid system, with yeast strain Y187 mated to PJ69-2A, and
then they were inoculated in QDO SD medium, when they grown up, glycerol
was added and quick freezed by liquid nitrogen and stored in -80C
At first, I thaw them and used a loop to streak them out on QDO plates,
they don't grow up, then I thaw them again and aliqioted 20ul and
spreaded on minus LT plates(less stringent I think), and just a few of
them grow up. 
Anything I can do to improve the recovery?
Thanks for your help!!!!!

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