Jose M Chaverri <jmc14009 at PEGASUS.CC.UCF.EDU> wrote:
> To whoever is out there doing in situ Hybridisation on drosophila embryos
> (whole mount), using asymetric PCR. I have problems with the signal (it
> is weak), and i have tried everything. I get good yields after PCR, but
> when I proceed with the in situ, the signal is not as strong as expected.
> Am i asking for too much?......
> sincerely yours
Which protocol are you using? Try playing around with the amount of
template DNA, labelled probe, hybridization temperature. Probes seem to
react differently, too. Most give good results with PCR, a few show
always better signals after labelling with random hexamers (at least in