yeast two hyb picture

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yeast two hyb picture

SL-3
Hi,

I am new doing yeast two hyb, and just got some results. My question is,
how to representing the Y2H result for a manuscript? I saw quite some
really nice pictures showing either colonies or streaks on plates, and
some of them have a duplicated one for LacZ assay. Can any one here give
me some detailed information on how to make this? Thank you so much.

SL
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Re: yeast two hyb picture

Philipp Pagel
SL <[hidden email]> wrote:
> I am new doing yeast two hyb, and just got some results. My question is,
> how to representing the Y2H result for a manuscript? I saw quite some
> really nice pictures showing either colonies or streaks on plates, and
> some of them have a duplicated one for LacZ assay.

You can buy a device we used to call "frog": It's a metal plate with a
handle that has 48 little metal stamps (~4mm diameter metal rods)
attached to it.  These metal stamps fit exactly into the wells of half a
96-well plate. So all you do is dissolve the colonies of interest in
e.g. 100-200ul of water in the 96well plate and stamp them onto a
selection plate. To sterilize the thing just dip it in ethanol and light
it.

I'm afraid I don't know what the correct name of the "frog" is. I would
just call the major lab suppliers and describe the thing to them - they
will figure it out...

The thing looks roughly like this


from the side:

            ______   Handle
           |      |
           |      |
    ======================  base plate
    I  I  I  I  I  I  I  I
   
               metal rods



from the bottom:

   ________________________
  |                        |
  | o  o  o  o  o  o  o  o |
  |                        |
  | o  o  o  o  o  o  o  o |
  |                        |
  | o  o  o  o  o  o  o  o |
  |                        |
  | o  o  o  o  o  o  o  o |
  |                        |
  | o  o  o  o  o  o  o  o |
  |                        |
  | o  o  o  o  o  o  o  o |
  |________________________|


cu
        Philipp

--
Dr. Philipp Pagel                          Tel. +49-8161-71 2131
Dept. of Genome Oriented Bioinformatics    Fax. +49-8161-71 2186
Technical University of Munich
http://mips.gsf.de/staff/pagel
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Re: yeast two hyb picture

Philipp Pagel
SL <[hidden email]> wrote:
> So it is basically suspend yeast colonies in water, and then spot them
> to a plate?

Yes.

> I might try my best to make it with a pipet

Maybe try a simple metal rod instead - that will give you nice round
spots.


cu
        Philipp

--
Dr. Philipp Pagel                          Tel. +49-8161-71 2131
Dept. of Genome Oriented Bioinformatics    Fax. +49-8161-71 2186
Technical University of Munich
http://mips.gsf.de/staff/pagel
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