16s metabarcoding of S. exigua hindgut
Total RNA was extracted following the TRItidy G manufacturers protocol and resuspended in RNase-free Milli-Q water. RNA concentration was quantified using a NanoDrop spectrophotometer, and RNA integrity was verified on a 1% agarose gel. One microgram of purified RNA was treated with DNase I (Thermo Fisher Scientific) according to the manufacturer's instructions and subsequently used for cDNA synthesis using the PrimeScript RT Reagent Kit (Takara Bio Inc., Kusatsu, Japan). cDNA was sent to Microomics Systems, for amplification of the V3V4 regions, library preparation, and Illumina MiSeq paired-end sequencing (2 x 300 bp; ~ 60,000-80,000 reads per sample). Amplification used primers SD-Bact-0341-b-S-17 (5-CCTACGGGNGGCWGCAG-3) and S-D-Bact-0785-a-A-21 (5-GACTACHVGGGTATCTAATCC-3)