5_Thyroid
Total RNA was extracted from isolated thyroid and lower jaw cartilage tissues from St. 46 tadpoles, as well as from fed and unfed tadpoles at Day 3, 6, 9 after feeding onset. Duplicate samples were prepared for each time point, except for Day 9, where only a single sample was obtained due to the limited number of tadpoles. The thyroid regions were dissected using a thin razor and fine forceps, and 20-40 pieces per stage were collected and homogenized in TRIzol LS reagent (#10296010; Thermo Fisher Scientific). RNA purification was performed using the RNA Clean & Concentrator-5 (R1014; Zymo Research, Irvine, CA, USA). Purified total RNA was outsourced for RNA-seq library preparation and sequencing to Takara Bio Inc. Libraries were generated using SMART-Seq v4 Ultra Low Input RNA Kit (Takara Bio) and sequenced on an Illumina NovaSeq 6000 platform, following the service providers protocols.