Illumina Genome Analyzer IIx sequencing; Meloidogyne incognita Egg Transcriptome
Egg samples were collected from tomato roots (Solanum esculentum cv. St Pierre) by incubation in 10% (v-v) Pectinex (Novozymes, Bagsvaerd, Denmark) and 5% (v-v) Celluclast BG (Realco, Louvain-la-neuve, Belgium) for 3 hours, respectively 10, 40 and 60 days after inoculation. Egg 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 resuspended 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 superior to 7). Reverse transcription was carried out using the Ovation pico WTA System (NuGEN Technologies, Inc, San Carlos, CA, USA).