[Zbrafish] About probe size for whole mount in situ

Lucas via zbrafish%40net.bio.net (by ilovequestions99 from gmail.com)
Wed Jan 11 21:13:19 EST 2012


Dear all,

Sorry to bother those who are not interested!

We just conducted whole-mount in situ hybridizations on zebrafish embryos (24-72 hpf) for hormone receptor genes using DIG-labeled probes that were ~1 to ~1.5 kb in length?  However we saw no staining for all 7 different receptors despite their strong expressions detected by PCR analyses. On the contrary, the probe for the ligands showed strong signals. 

We are wondering if this is because of the dispersed expressions of the hormone receptors,or there are some other reasons that we simply don't know? And do you think that this method could be more efficient when using a smaller sized probe?

Please share your thoughts!

Thanks

Olivia
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