in-situ hybridization

Emma Rushton rushton at
Mon Oct 27 15:28:11 EST 1997

Sounds to me as though your proteinase K is too strong, or you are leaving
it on too long.  Try a few different dilutions to titrate it.  Have the
glycine to block it ready at hand and put it on the moment the incubation
ends - I remove prot K BEFORE the incubation is over and add glycine as
soon as my timer goes off.

Hope this helps, Emma

In article <Pine.A41.3.96.971024181656.171304A-100000 at>,
siau min fung <siauminf at> wrote:

> hi!!!!! 
> I am doing mRNA in-situ hybridization on fly embryo right now!!!! I have
> repeat twice and I have problem with embryo aggregate together and when I
> pipet up and down to separate them, the embryo break into pieces. 
> Does anybody has any idea why is that???? The protocol that I followed is
> the typical in-situ hybridization protocol to whole embryo.
> any suggestions will be highly appreciated!!!!!!!!
> Thank you very much!!!!!!
> SiauMin
> siauminf at

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