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.
Vanholme
Bartel
B
Vakgroep Moleculaire Biotechnologie, Faculteit Landbouwkundige en Toegepaste Biologische Wetenschappen, Universiteit Gent, Coupure links 653, 9000 Gent, Belgium.
De Meutter
Jan
J
Tytgat
Tom
T
Gheysen
Greetje D C
GD
Vanhoutte
Isabelle
I
Gheysen
Godelieve D R
GD
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