RNAseq of Helmeted honeyeater
5 L of mRNA was used to prepare the library using the NEBNext Ultra RNA Library Prep Kit (Illumina). In adaptor ligation step, the NEBNext adaptor was diluted 25-fold in dilution buffer. The PCR enrichment of adaptor-ligated RNA cycling conditions, denaturation, and annealing/extension cycle steps were repeated with a total of 15 cycles. Each library was pooled to equal molar concentrations, enzymatic treated, denatured and normalized to 2 nM.