I am designing the experiments for my PhD thesis and have no experience on ISH. I would like to have an expression pattern of a protein in the CNS of 3 dpf zebrafish larvae. The target transcript is about 1.5 kb and for simplicity I intend to order the synthesis of the oligo template from an external company. Any recommendations on this?
Rebecca Ann Wingert
Yes, the Thisse protocol is very effective on 3 dpf zebrafish embryos.
The size of the template you are intending to use should enable specific labeling and adequate infiltration of a riboprobe; if your signal is weak one method to trouble shoot this is to fragment your probe into shorter lengths to improve penetration into the target tissue(s).
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