An improved method for whole-mount in situ hybridization of Heterodera schachtii juveniles.
An optimized protocol is presented to visualize gene expression in the sedentary beet cyst nematode, Heterodera schachtii, by whole-mount in situ hybridization. Two different probes were used for genes with known expression pattern in other nematodes. Vacuum infiltration of the fixative significantly increased its efficiency and resulted in a nicely preserved morphology. Additional modifications were introduced to simplify and standardize the process.
Vakgroep Moleculaire Biotechnologie, Faculteit Landbouwkundige en Toegepaste Biologische Wetenschappen, Universiteit Gent, Coupure links 653, 9000 Gent, Belgium.
Bartel
B
Vanholme
Jan
J
De Meutter
Tom
T
Tytgat
Greetje D C
GD
Gheysen
Isabelle
I
Vanhoutte
Godelieve D R
GD
Gheysen
D000818:Animals; D015342:DNA Probes; D018507:Gene Expression Regulation, Developmental; D017403:In Situ Hybridization / Q000379:methods* / Q000592:standards; D010935:Plant Diseases / Q000469:parasitology; D016133:Polymerase Chain Reaction / Q000379:methods; D018510:RNA, Helminth / Q000032:analysis; D012680:Sensitivity and Specificity; D014431:Tylenchoidea / Q000235:genetics* / Q000254:growth & development / Q000648:ultrastructure
D015342:DNA Probes; D018510:RNA, Helminth