Small RNA-Seq of Dermatophagoides pteronyssinus: Mixed stages whole body
Dermatophagoides pteronyssinus were cultured at 25 C in a small air-filtered room at 70-75% relative humidity for 50 days. Total RNA was isolated from the homogenate with Trizol followed by PureLink RNA Mini Kit (Thermo Fisher Scientific). The molecules in the range of 18-44nt were enriched by polyacrylamide gel electrophoresis. These small RNAs were ligated with adapters and were used as templates for cDNA library. The quantity and quality of the RNA sample were measured with a NanoDrop 2000 Spectrophotometer (Thermo Fisher Scientific) and Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA). Next-generation sequencing was performed using Illumina HiSeq 2000 to produce pair-end reads at 90bp per end.