WormBase Tree Display for RNAi: WBRNAi00106289

WBRNAi00106289 Homol: Homol_homol: R13G10:RNAi
  Sequence_info: PCR_product: sjj_R13G10.1
  Experiment: Treatment: For DPY-27 RNAi, the bacterial strain from the Ahringer RNAi library containing dpy-27 RNAi plasmid and control plasmid were grown and induced in 100 ml LB media for 3 hours with 0.1 mM ITPG, concentrated 10-fold and seeded onto a 10 cm NGM plate supplemented with ampicillin, tetracycline and 1mM IPTG. Synchronized L1s were placed on seeded plates and collected at 24 hours (L3 larvae) or grown to gravid adults and embryos were collected by bleaching (mixed stage embryos). At collection time, all samples were washed 3 times in M9 buffer and stored in ten volumes of Trizol (Invitrogen) at -80C. 2-3 collections were pooled for one biological replicate.
    Delivered_by: Bacterial_feeding
  Inhibits: Predicted_gene: R13G10.1 Inferred_automatically: RNAi_primary
    Gene: WBGene00001086 Inferred_automatically: RNAi_primary
    Transcript: R13G10.1.1 Inferred_automatically: RNAi_primary
  Species: Caenorhabditis elegans
  Reference: WBPaper00048953
  Phenotype: WBPhenotype:0000718 Remark: "To specifically study DCC mediated X repression, we analyzed gene expression changes in hermaphrodites mutant for dpy-27 or upon dpy-27 RNAi knockdown. As mutants and RNAi treated worms showed more variability in staging, we mainly used 'mixed stage embryos' isolated by bleaching gravid adults. Mixed stage embryos contained 100-300 cells (S4A Fig). We used dpy-27 RNAi in mixed embryos and L3, because of the difficulty collecting dpy-27(y56) null mutant. Western blot analyses showed ~70% and ~40% knockdown of DPY-27 in embryos and L3s, respectively (S4B Fig). Mutation (Fig 3D) or depletion (Fig 3E) of dpy-27 caused significant X chromosome derepression in early and mixed-stage embryos, L1 and L3 worms."
  Remark: dpy-27 RNAi
  Method: RNAi