Polypedates megacephalus Skin Transcriptome
Total RNA was extracted from the skin of frogs using Trizol reagent (Invitrogen, Carlsbad, CA, USA) and purified using RNeasy Animal Mini Kit (Qiagen, Valencia, CA, USA) according to the manufacturer's instructions. mRNA was isolated from the total RNA using poly-T oligo-attached magnetic beads (Invitrogen, Carlsbad, CA, USA) and was fragmented into short fragments using divalent cations under elevated temperature. After that, the short fragments were connected to adapters. Suitable fragments were selected as templates for PCR amplification and abundance normalization. During the QC steps, an Agilent 2100 Bioanaylzer (Agilent, Santa Clara, CA, USA) and an ABI StepOnePlus Real-Time PCR System (ABI, Waltham, MA, USA) were used in quantification and qualification of the sample library. At last, the library was sequenced using Illumina HiSe 2000 (Illumina, San Diego, CA, USA).