I am currently trying to label a message on 0-4hr CS embryos using a DIG
labelled riboprobe. My problem is that, despite the fact that my
positive control works fine and that my transcript is abundant, I
repeatedly see no staining. Can anyone please suggest avenues by which I
can try to improve my protocol with respect to the size of riboprobe
(with or without alkaline hydrolysis), time of prehyb and probing,
temperature, fixation, etc., as I am making little progress in
elucidating the expression pattern of my transcript.
Many thanks for your help.
E-mail me directly or to the group.
mjcann at med.cornell.edu