[Histonet] Re: chicken in situ hybridization

[dengqiaolin deng]

   Hi, Teri:

   Thank  you  for  your  points. In my protocol, I treat the slides 5min
   with  proteinase  K,  but  the my chicken embryo is E4. Maybe I should
   shorten  the  time.  Also,  I want to ask how you do the hybridization
   step.  In  our lab, we drop 100ul hybridization solution with probe on
   the slide and then put a coverslip on it. The day after, we remove the
   coverslip by dipping the slides in the ssc solution. I find it is also
   a reason for losing the structure. Do you have other way to do it?

   Best regards

   Qiaolin
   >From: "Johnson, Teri" <TJJ <@t> Stowers-Institute.org>
   >To: <histonet <@t> lists.utsouthwestern.edu>
   >Subject: [Histonet] Re: chicken in situ hybridization
   >Date: Mon, 24 May 2004 08:40:16 -0500
   >
   >Qiaolin,
   >
   >Are  you  using  proteinase K enzyme digestion in your protocol?  You
   may
   >not  need  it  on  very young chick embryo cryosections.  We've found
   that
   >we don't need it most times on E2 whole mount ISH.
   >
   >Teri Johnson
   >Managing Director Histology Facility
   >Stowers Institute for Medical Research
   >1000 E. 50th St.
   >Kansas City, Missouri  64110
   >tjj <@t> stowers-institute.org
   >
   >
   >
   >
   >Date: Sun, 23 May 2004 15:17:41 +0000
   >From: "dengqiaolin deng" <qiaolin_deng78 <@t> hotmail.com>
   >Subject: [Histonet] chicken in situ hybridization
   >To: histonet <@t> lists.utsouthwestern.edu
   >Message-ID: <BAY7-F92wWJv3zOdnjQ00034afa <@t> hotmail.com>
   >Content-Type: text/plain
   >
   >
   >    Dear histonetter:
   >
   >            Is  there anyone having done in situ hybridization for
   >chicken
   >    cryosection?  One  kind  of  problems  I  have  met  is  that the
   structure
   >is
   >    easy  to  be crashed and disattached from the slide although I am
   >very
   >    careful  with  every  washing  steps.  But  for the mouse
   >cryosection,
   >    everything  is  fine.  Is there any improvement that I can do to
   >avoid
   >    it?
   >          Thank you for any suugestions in advance.
   >
   >    Qiaolin Deng
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References

   1. http://g.msn.com/8HMBEN/2737??PS=47575