Skarmeta Lab, Centro Andaluz de Biologia del Desarrollo
Description:
10 zebrafish embryos were manually dechorionated and disrupted in 500 µl of Ginzburg Fish Ringers (55 mM NaCl, 1.8 mM KCl, 1.25 mM NaHCO3). After washing with cold PBS, 75.000 cells were lysed (lysis buffer: 10 µM Tris pH7.4, 10 µM NaCl, 3 µM MgCl2, 0.1% IGEPAL) and incubated for 30 min at 37ºC with the TDE1 enzyme. The sample was then purified with Qiagen Minelute kit, and a PCR was performed with 13 cycles using Ad1F and Ad2.1R primers and KAPA HiFi hotstart enzyme (Kapa Biosystems).