Dermatophagoides pteronyssinus metatranscriptome
Lyophilized total RNA was first resuspended in 25uL RNase/DNAse free water and the concentration of total RNA was determined. 1ug total RNA was used for rRNA removal by using Ribo-Zero Plus rRNA Depletion Kit (Illumina). Libraries for rRNA depleted samples were prepared using the KAPA mRNA HyperPrep Kits (Roche) by following the manufacturer's instructions. Following the library preparation, the final concentration of the libraries was measured using the Qubit dsDNA HS Assay Kit (Life Technologies), and the average library size was determined using the Agilent 2100 Bioanalyzer (Agilent Technologies). The libraries were then pooled in equimolar ratios of 0.6nM, and sequenced paired end for 300 cycles using the NovaSeq 6000 system (Illumina).