[Histonet] RE:Re: chicken in situ hybridization
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?
>From: "Johnson, Teri"
>To: "dengqiaolin deng"
>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
>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.)
> -----Original Message-----
> From: dengqiaolin deng [mailto:firstname.lastname@example.org]
> Sent: Monday, May 24, 2004 10:22 AM
> To: Johnson, Teri
> Cc: email@example.com
> 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
>shorten the time. Also, I want to ask how you do the hybridization
>In our lab, we drop 100ul hybridization solution with probe on the
>and then put a coverslip on it. The day after, we remove the
>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
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