in-situ hybridization

[Emma Rushton]

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 tigger.cc.uic.edu>,
siau min fung <siauminf at uic.edu> 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 uic.edu