RNA-Seq of Lucilia cuprina: whole body tissue
Illumina RNA library construction and sequencing were performed by the North Carolina State Genomic Sciences Laboratory (Raleigh, NC, USA). Samples were enriched for mRNA using oligo-dT beads provided in the NEBNExt Poly(A) mRNA Magnetic Isolation Module (New England Biolabs, USA). cDNA libraries for Illumina sequencing were constructed using the NEBNext Ultra II Directional RNA Library Prep Kit (NEB) and NEBNext Mulitplex Oligos for Illumina (NEB) using the manufacturer-specified protocol. Samples were selected for a final library size (adapters included) of 400-550 bp using sequential AMPure XP bead isolation (Beckman Coulter, USA). The final quantified libraries were pooled in equimolar amounts for clustering and sequencing on an Illumina NovaSeq 6000 DNA sequencer, utilizing one lane of an S4 150x2 PE sequencing reagent kit (Illumina, USA), generating approximately 50M reads per sample.