[Histonet] RE:Re: chicken in situ hybridization

[dengqiaolin deng]

   Hi, Teri:

        Thank  you  very  much  for  your  further suggestions. As to the
   slides,  we  use  SuperFrost  Plus  slides which is ready to use. So I
   don't  do  anything  to  the  slides.  Before starting in situ, I only
   air-dry  the  slides  briefly.  Actully,  I  didn't  realizae it is an
   important step before you point it out. How long do you usually do?

   Qiaolin
   >From: "Johnson, Teri" <TJJ <@t> Stowers-Institute.org>
   >To: "dengqiaolin deng" <qiaolin_deng78 <@t> hotmail.com>
   >CC: <histonet <@t> pathology.swmed.edu>
   >Subject: RE: [Histonet] Re: chicken in situ hybridization
   >Date: Mon, 24 May 2004 10:38:26 -0500
   >
   >Try  lowering  the  concentration  of  proteinase  K  rather than the
   time.  If
   >you have an additional slide, try it with no pretreatment and see if
   >that makes a difference.  We remove the coverslip the same way, by
   >dipping it into the SSC.  If that isn't working for you, try just
   >leaving the slides in SSC without the dipping motion and letting the
   >coverslips just fall off on their own.  How long do you let your
   >cryosections air-dry before doing your protocol?  And do you use any
   >adhesive on your slide (+ charged, poly-l-lysine, etc.)
   >
   >Teri
   >
   > -----Original Message-----
   > From: dengqiaolin deng [mailto:qiaolin_deng78 <@t> hotmail.com]
   > Sent: Monday, May 24, 2004 10:22 AM
   > To: Johnson, Teri
   > Cc: histonet <@t> lists.utsouthwestern.edu
   > Subject: RE: [Histonet] Re: chicken in situ hybridization
   >
   >
   >
   > 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
   >
   >
   >
   >
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