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Yeast Forum Discuss research on yeast the model organism. Post questions and topics on the molecular biology and genetics of Saccharomyces cerevisiae and other yeast species.


yeast two hybrid

yeast two hybrid - Yeast Forum

yeast two hybrid - Discuss research on yeast the model organism. Post questions and topics on the molecular biology and genetics of Saccharomyces cerevisiae and other yeast species.


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Old 08-27-2003, 06:02 PM
Alexandra Blak
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Default yeast two hybrid



Dear yeasters,

after trying a YTH screen with a pretransformed library, that didn`t
work so well, I created my own cDNA library by using Clontech`s MM
library construction and screening kit and I have to say that the
screen worked really nice (after having a lot of troeble with every
single step)! But now my problem is: I got around 500 colonies and
although I theoretically know what to do now, like PCR, extracting the
bands out of a gel, cutting them with Alu or Hae; colony hybridisation
(with what?), I am really worried about that number - 500. Does anyone
of you know a better and faster way of sorting out false positives? I
restreaked my colonies and most of them still grow. alpha-Gal assay
didn`t work on plates - they are all getting blue and for beta Gal
assay I made the same experience. Did somebody also use the MM library
construction kit? I would be very very thankful for every little
hint. Thanks a lot in advance. Alexandra

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