title: Hsch_IRS
organism: Heterodera schachtii
description: Protocols: H. schachtii cysts (Woensdrecht population from IRS, the Netherlands) were separated from sand of infected Brassica oleracea plants as previously described (Baum et al., 2000). J2s were concentrated, and sucrose centrifugation was used to purify the nematode suspension (Jenkins, 1964). After multiple rounds of washing the purified nematode suspension in 0.1 M NaCl, nematodes were resuspended in sterilized MQ water. Juveniles were lysed in a standard nematode lysis buffer with proteinase K and beta-mercaptoethanol at 60°C for 1 hr as described in Holterman et al. (2006). The lysate was mixed with an equal volume of phenol/chloroform/isoamyl alcohol (25:24:1) (pH 8.0) following a standard DNA purification procedure, and finally, DNA was precipitated with isopropanol. DNA fragments of <10 kb were depleted using a short read eliminator kit (Circulomics SS-100-121-01). The sample was send to USEQ for library preparation. Library preparation was done using the Truseq DNA nano kit at USEQ.