Illumina HiSeq 2000 paired end sequencing; Meloidogyne javanica Egg and J2 Transcriptome
Eggs and J2 were collected from tomato roots (Solanum esculentum cv. St Pierre) by incubation in 10% (v for v) Pectinex (Novozymes, Bagsvaerd, Denmark) and 5% (v for v) Celluclast BG (Realco, Louvain-la-neuve, Belgium) for 3 hours, 60 days after inoculation. Samples were purified from root debris by sucrose gradient centrifugations.
RNA isolation using TRIzol Reagents (Invitrogen, Carlsbad, CA, USA) was done according to protocol available from Invitrogen and re-suspended in 10 ul RNase-free water. Purity and concentration of the RNA was determined on a 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA) and cDNA was only produced from high RNA quality (RIN greater than 7). Reverse transcription was carried out using the Ovation pico WTA System (NuGEN Technologies, Inc, San Carlos, CA, USA).