smallRNA-Seq of Rhodnius prolixus: ovarioles (previtellogenic stage)
Previtellogenic stages and mature unfertilized eggs (~30 eggs per biological replicate) were dissected in ice cold 1X PBS from adult females two weeks after the blood meal. Total RNA was extracted with Trizol Reagents (Life Technologies) as per manufacturer instructions and treated with turbo DNA-free kit (Ambion). 20ug of total RNA was separated on 15% TBE-Urea gels and small RNAs ranging in size 18-30nt were selected as previously described. Library preparation and sequencing were performed at the Max Planck Institute Sequencing Facility with Illumina protocols and NextSeq sequencing platforms.
SUB9529885
RPPVS