title: Metatranscriptome of lab-reared Spodoptera frugiperda larvae guts from Tucumán province (Argentina)
design: Fifth instar S. frugiperda larvae reared for 9 generations in the laboratory, were sacrificed by exposure to low temperatures, surface sterilised before dissection, and the digestive tracts were extracted using sterile entomological forceps. For the extraction of total RNA, intestines were stored in sterile 1X Phosphate Buffered Saline (PBS) at -80oC for later processing.  Two digestive tracts were pooled and ground in Tri-Reagent (Molecular Research Center Inc., Cincinnati, OH) with a sterile Teflon pestle, and total RNA was immediately extracted according to the manufacturer’s instructions. Total RNA was submitted to a modified one-step reverse transcription and polymerase chain reaction (RT-PCR) sequence-independent amplification procedure. Briefly, RevertAid™ Premium (Fermentas, Vilnius, Lithuania) was used for a first-strand reverse transcription which was initiated with a random octamer linked to a specific primer sequence (primer 1: 5’-GTT TCC CAG TAG GTC TCN NNN NNN N-3’). cDNA was then amplified with the Expand Long Template PCR Enzyme mix (Roche) using the above primer and a primer targeting the specific primer sequence (primer 2: 5’-CGC CGT TTC CCA GTA GGT CTC-3’). Samples were submitted for high-throughput pyrosequencing (Macrogen Inc., Korea).