E. coqui stg 9 whole tail RNAs following T3 treatment
E. coqui embryos development stage 9-10 were exposed 24 hours with 10 nM T3. The whole tail was dissected and lysed with tissue lyser II (QIAGEN) and a stainless-steel ball (INOX AISI 304 grade 100 AFBMA) in 500 L RNAble (Eurobio). Tubes were transferred on ice and 100 L chloroform added. Samples were then centrifugated 12 000 g at 4C during 15 min. Next, 175 L of AquaPhenol (Q-Biogene: AquaPH01) and 100 L of chloroform were added to 350 L of supernatant. Following centrifugation 12 000 g at 4C during 15 min, RNAs from 250 L of the supernatant were recovered with RNeasy Mini Kit (Qiagen: 74104). Sequencing libraries were prepared with TruSeq RNA Sample Prep v2 kit.