RNA Detection

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  1. Hearts are dissected from adult zebrafish using iridectomy
    scissors and Dumont #5 fine-tip forceps and placed directly
    into 1.5 mL centrifuge tube containing 1 mL of tissue fixative.
    Fix hearts at room temperature for 1 h. Alternatively, keep
    hearts in fixative overnight at 4C(seeNote 5).


Fig. 2A humidified chamber for ISH incubations. Serological pipettes are cut to the appropriate length and
attached to the bottom surface of the Tupperware with laboratory tape. The distance between the each set of
pipettes is adjusted to accommodate the width of the slide. Water soaked paper towels are placed inside the
chamber to create a moist environment for incubations


Fig. 3The adult zebrafish heart. The zebrafish heart is a two-chambered organ. When extracting the organ for
histology, it is important to keep the outflow tract, atrium, and ventricle as one unit to serve as landmarks in
histology


ISH Detection of MicroRNA and Target Gene mRNAs 201
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