[Histonet] In situ hybridization

[sebres]

Here is an excellent web site on ISH:
http://intramural.nimh.nih.gov/lcmr/snge/Protocols/ISHH/ISHH.html
----- Original Message ----- 
From: "Jimmy Lao" <lao_ji <@t> yahoo.com>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Wednesday, November 26, 2003 12:20 PM
Subject: Re: [Histonet] In situ hybridization


> Susan Wells,
> This is exactly a set of four books.
> ISBN: 047150338x
> I searched half.com, there are new and used book of
> this item there. new one cost $647.30, the used one
> $100 but only volumn 1&2.
> I couldn't recommend our Lab wibsite due to under
> construction. but for in situ protocol, you can  
> visit:http://www.rodentia.com/wmc/docs/Big_In_Situ.html
> I found it is similar to the one I used.
> Hope this help.
> Zhimin Lao
> 
> --- Susan Q Wells <susan.wells <@t> bms.com> wrote:
> > A few days ago I saw a post from you regarding a
> > book I'm having
> > trouble finding.
> > Current protocols in molecular biology
> > volume2.edited by
> > Frederick Ausubel et al.Printed by John Wiley&sons
> > inc.Could you
> > check the book again.I cannot find anything on this
> > book and I am
> > intersetd in ordering it.Perhaps Frederick Ausubel
> > wrote it or I
> > have the title wrong?
> > Thanks,
> > Susan Wells HT,QIHC(ASCP)
> > 
> > Jimmy Lao wrote:
> > 
> > > Hi, Carlene Zindi,
> > > It seems you are going to do In situ hbridization
> > on
> > > slides. I would like to answer your question based
> > on
> > > this point with my experience.
> > > 1. We used dig conjugated RNA probes like fgf8,
> > otx2,
> > > Shh and so on for mouse embryo and the brain,
> > limbs
> > > from bigger than E12.5 mouse.  They work fine.
> > > 2. The fix method is that 4% paraformaldehyde in
> > PBS
> > > at 4 C overnight with gentl nutation. Wash in PBS
> > 5
> > > min at room temperature for two times. embed in
> > OCT
> > > crystat section 10-14um thickness.
> > > 3. About 1Kb anti-sense probe is reasonable, and
> > it is
> > > better using 3' UTR(untranslation region) due to
> > not
> > > much conserved.
> > > Hope this help.
> > > Zhimin Lao
> > >
> > > --- "Carlene L. Zindl" <clzindl <@t> artsci.wustl.edu>
> > > wrote:
> > > > Hello,
> > > >
> > > > Does anyone have experience with non-radioactive
> > in
> > > > situ hybridization of
> > > > frozen mouse tissue??  If so, I would appreciate
> > > > advice on details such
> > > > as: 1) is biotin or dig better? 2) what is the
> > best
> > > > way to fix this
> > > > tissue?  3) what is the optimal size RNA probe?
> > > >
> > > > Thank you for any pertinent information.
> > > > Sincerely,
> > > > Carlene Zindl
> > > > University of Alabama at Birmingham
> > > > clzindl <@t> artsci.wustl.edu -OR-  clzindl <@t> uab.edu
> > > >
> > > >
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> 
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