Embryonic Microinjection and In Situ Hybridization
Purified DNA construct was diluted to 50 ng/µl with water, and injected into cell body at 1-cell stage. Developing embryos were sorted and cultured at 28.5 °C until the appropriate stage. To prepare embryos for in situ hybridization, we added phenylthiourea (Sigma-Aldrich, Cat. No. P-7629) at 0.003%, whereas a quarter dose was used to allow them to be only weakly pigmented when we needed to detect fluorescent signals in melanocytes after antibody staining.
Phenylthiourea (PTU) concentration: 0.003% for full pigmentation and a quarter dose for weak pigmentation
controls
Positive control: Not specified
Negative control: Not specified
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