RNA-Seq of Colorado potato beetle S16
The liquid phase of the whole Leptinotarsa decemlineata prepupa homogenate was collected and filtered through a Millex PVDF syringe with a Millex-HV filter (0.45 nm) and concentrated to 200 l using an Amicon Ultra 4 50K Millipore centrifuge filter. A 100 l aliquot was incubated with benzonase. Viral RNA was extracted with Trizol using glycogen as a co-precipitant. The dissolved precipitate was used to obtain cDNA with SISPA method. The resulting ds-cDNA was purified with AMPure beads. DNA concentration was measured with a Qubit 3.0 using the Qubit dsDNA HS Assay Kit. Y adapters were used to prepare the sequencing library. Sequencing was performed using the Illumina iSeq 100 platform using NEBNext Ultra II FS DNA Library Prep Kit for Illumina.