title: GSM8653011: Control Healthy 2 6h; Galleria mellonella; RNA-Seq
library_construction_protocol: RNA isolated at 6, 30, 72 and 168 hours post-infection. The contents of three larvae were pooled, flash-frozen with liquid nitrogen and mechanically crushed using a pestle and mortar. The resulting powder was suspended in RLT buffer (supplemented with 1% β-Mercaptoethanol), provided in the RNeasy Mini Kit (Qiagen, Germany). The samples were incubated at 57⁰C for 3 minutes before proceeding according to the manufacturer's instructions. LQ-ssCAGE (low quantity single strand CAGE) started by Reverse Transcription (RT) of 200 ng of total RNA (*measured by the RNA provider) to generate 5'-end of the capped RNA/first-strand cDNA hybrids. The RNA/cDNA hybrids were cap-trapped and adapted with illumine sequencer-compatible barcode-tagged adaptors (PMID: 34382184). No PCR amplification was performed during the LQ-ssCAGE library preparation. The amount of resulting libraries were checked by qPCR using KAPA Library Quantification Kits (Kapa Biosystems). LQ-ssCAGE library sequenced using Illumina HiSeq2500 platform in Rapid mode and 50 base read length.