RNA-seq of Spodoptera frugiperda larvae strains (SfC and SfR) and hybrids (CR and RC)
RNA-seq of Spodoptera frugiperda larvae strains (SfC and SfR) and hybrids (CR and RC)
Rearing of 30 larvae per genotype has been performed in individual cups filled with artificial diet. 1 day after molting of 4th instar larvae (L4), individual caterpillars were placed in a 2 mL Eppendorf tube with RLT Plus buffer from AllPrepr DNA/RNA kit  (Qiagen). Larvae were then immediately disrupted and homogenized with a TissueLyser II (Qiagen #85300) with the aid of a 5mm metal bead. No dissection has been performed since whole individual larvae have been used. Larvae were reared on individual cups with artificial diet. 3Ã  larvae per genotype, in triplicate have been reared in parallel in the same conditions within an incubator of the DGIMI quarantine facility for insects (PIQ). No treatment has been performed. The experimental design consists of parralel rearing of four different genotypes. Two parental strains CC and RR, and their reciprocal hybrids CR and RC (first letter is the mother type). Homogenate was then processed with the AllPrep DNA/RNA kit (Qiagen # 80204) according to manufacturer's protocol to purify bot DNA and RNA from the same sample. DNA contaminants from RNA sample was removed by TURBO-DNA-free kit (Invitrogen # AM1907). Amount and purity of DNA was measured by Nanodrop to match the requirements of the sequencing company (Novogene). Libraries have been performed on total RNA by Novogene. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection.