Xiphinema index transcriptome. ANU

Mixed developmental stages of X. index, reared in greenhouse on fig plants, were collected and pooled together for RNA extraction and cDNA library preparation. To maximize the number of expressed genes, we produced X. index specimens both in standard and stress conditions.Stressed X. index were obtained by soaking for 1 h in 0.2 M NaCl, for 30 min in 1% EtOH, for 1 h at 40°C or for 1h in 0.1% acetic acid. Nematodes from stress treatments were pooled before RNA extraction.Total RNA was extracted using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) and reverse transcription was carried out using the Ovation pico WTA System (NuGEN Technologies, Inc, San Carlos, CA, USA).The two libraries, non-stressed and stressed, were sequenced on an Illumina GAIIx sequencer to produce 76 nt single end reads.