Sugarcane giant borer transcriptome analysis and identification of genes related to digestion
Total RNA was extracted separately from each insect stage, eggs, early-stage larvae, late-stage larvae, prepupae, pupae, male and female adults using Trizol Reagent (Invitrogen, Life Technologies), according to the manufacturer’s protocol. RNA was treated with RNAse-free DNase I (Ambion, Life Technologies) at 37 oC for 30 minutes, according to the manufacturer’s protocol. The normalized transcripts were submitted to a half-plate run using the 454 pyrosequencing, GS FLX Titanium technology, according to protocols provided by manufacturer (Roche 454 Life Sciences).