title: Illumina Genome Analyzer IIx paired end sequencing; Longitudinal and Diurnal Experiments of M. hapla and M. truncatula
design: Longitudinal and Diurnal Experiments of M. hapla and M. truncatula
library_construction_protocol: The experiments were conducted in greenhouse conditions. Meloidogyne hapla in all experiments were maintained on the root system of Tomato (cv. Rutgers). An egg mass of M. hapla isolated by the standard bleach procedure was purified on a 40% sucrose density gradient as previously described (Byrd et al. 1996). For sterile M. hapla egg hatches, hatcheries were prepared with mesh-based sieves in a shallow bowl. The collection of viable infective J2 was obtained after three days of hatching. Host plant, Medicago truncatula, was synchronously inoculated with M. hapla J2. The acquisition of homogenous mixture J2 was supported by collecting over three days after egg isolation and hatching, which diluted the potential effects of post-hatch age of J2 on larval penetration variance.  RKN and host plant materials were harvested at indicated time points and immediately frozen in liquid nitrogen. RNA isolation procedure was followed by the Dynabeads® RNA DirectTM Purifciation Kit protocol. The RNA quality and concentration were checked on Bioanalyzer 2100 (Agilent, Santa Clara, CA, USA) and RNA Pico Chip respectively. Libraries were prepared using a TruSeq RNA library prep kit v1 (Illumina, San Diego, CA, USA).