[Histonet] Re: chicken in situ hybridization

[Johnson, Teri]

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