DNA-seq of 28 populations of Meloidogyne hapla
DNA-seq of 28 populations of Meloidogyne hapla
The nematodes were propagated on tomato cv. Money Maker. Fresh eggs were harvested 10 weeks after the nematodes were inoculated. Eggs were obtained by treating roots with 0.05% (v/v) NaOCl for three minutes, following by rinsing with tap water. After further rinsing, eggs were hatched on a 25 µm sieve. Hatched J2 were collected by centrifugation on a 70% sucrose column. The purified J2s were lysed with nematode lysis buffer with proteinase-K and beta-mercaptoethanol. This was done in a ThermoCycler at 60 ºC for 1 h with continuous shaking at 650 RPM. The lysate was then mixed with an equal volume of phenol:chloroform:isoamyl alcohol (25:24:1) (pH 8.0) following a standard DNA purification procedure, followed by precipitation with isopropanol. Afterwards, the DNA pellet was washed with 70% ethanol for two times, before finally resuspended with 10 mM Tris-HCL (pH 8.0). DNA library preparation and sequencing was done in Utrecht sequencing facility (USEQ). Library preparation was done using the Illumina TruSeq protocol.