Traditional small-RNA-seq after GFP-dsRNA injection to actin::GFP anemones 24-hours old gastrula replicate1
Nematostella actin::GFP zygotes were injected with dsRNA carrying GFP sequence then frozen in liquid nitrogen after 24 hours. RNA was extracted with Tri-reagent (Merck Millipore) according to manufacturer's protocol. Total RNAs were separated by 10% Urea-PAGE gel and 15-50nt small RNAs were recovered from the gel. The cDNA libraries were prepared with NEBNext Small RNA Library Prep Set for Illumina kit (New England Biolabs).