lungfish blastema transcriptome and differential gene expression analysis of blastema versus non-regenerating fin
Total RNA extraction from blastema or non-regenerating fin tissue for transcriptome or qPCR was achieved using TRIzol® Reagent (Life Technologies, Cat. 15596-026) according to the manufacturer’s protocol. RNA samples were further purified using RNeasy® Mini Kit (Quiagen, Cat. 74104) and treated with DNaseI (Quiagen, Cat. 74254), according to the manufacturer’s protocol. RNA (0.5 μg) was reverse transcribed using the SuperScript III First-Strand Synthesis SuperMix ( Life Technologies, Cat. 18080-400). Transcripts abundance was based on six independent Illumina 50 bp single-end runs, three runs from three non-regenerating fin libraries and three from blastema tissue.