RNA-seq from hypoxia pathway C. elegans mutants hif-1 (ia4) and vhl-1 (ok161)
RNA-seq from hypoxia pathway C. elegans mutants hif-1 (ia4) and vhl-1 (ok161)
Synchronized worms were grown at basal conditions (20Â°C) and collected at their day 1 adult stage in Trizol (Invitrogen). Total RNA was extracted using RNeasy Mini spin column (QIAGEN). Concentration and purity of the RNA was measured by NanoDrop. polyA+ mRNA was isolated from total RNA with NEBNext Poly(A) mRNA Magnetics Isolation Module (New England Biolabs). RNA-Seq libraries were prepared with NEBNext Ultra Directional RNA Library Prep Kit for Illumina (New England Biolabs). Capillary electrophoresis (Agilent Bioanalyser and Agilent Tapestation) was used to assess quality and quantity at all steps. Libraries were quantified by fluorimeter, immobilized and processed onto a flow cell with a cBot (Illumina).