In situ Hybridization of Catshark Jaws

Antisense RNA digoxigenin-UTP probes were transcribed using SP6 or T7 RNA polymerases (Roche), according to the orientation of the insert in the plasmid. In situ hybridizations were performed on dissected catshark lower jaws and dissected tails as previously described [41 (link)] with proteinase K treatments (10 μg/ml) as in [4 (link)]. The color detection step was performed using the NBT-BCIP reaction (Roche). Samples were post-fixed in 4 % PFA after whole mount in situ hybridization, then cleared and stored in glycerol at 4 °C until photographed.

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Debiais-Thibaud M., Chiori R., Enault S., Oulion S., Germon I., Martinand-Mari C., Casane D, & Borday-Birraux V. (2015). Tooth and scale morphogenesis in shark: an alternative process to the mammalian enamel knot system. BMC Evolutionary Biology, 15, 292.

Publication 2015

SP6 or T7 RNA polymerases

Digoxigenin Glycerol In situ hybridization Lower jaws Plasmid Proteinase k Rna probes T7 rna polymerases Tails

Corresponding Organization :

Other organizations : Institut des Sciences de l'Evolution de Montpellier, Évolution, Génomes, Comportement, Écologie

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Variable analysis

independent variables

Orientation of the insert in the plasmid

dependent variables

Gene expression patterns observed through in situ hybridization

control variables

Proteinase K treatments (10 μg/ml)
NBT-BCIP reaction for color detection

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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