WormBase Tree Display for RNAi: WBRNAi00103124

WBRNAi00103124 Homol: Homol_homol: F46E10:RNAi
  Sequence_info: PCR_product: sjj_F46E10.10
  Experiment: Laboratory: MR
    Date: 13 Jul 2012 00:00:00
    Genotype: daf-2(e1370); aak-1(tm1944); aak-2(ok524)
    Treatment: In brief, L3-L4 stage hermaphrodites were transferred onto regular plates seeded with individual double-stranded RNA-expressing bacterial clones. The animals were transferred to a new set of seeded plates after 1 day of incubation at 15 degrees Celsius, and phenotypes were scored for the F2 generation. Synchronized L1 larvae were grown at 25 degrees Celsius until they formed dauer. Dauer larvae were then collected and washed in PBS buffer for 3 times before the weight of the worm pellet was measured. The worms were resuspended 1:2 in PBS buffer supplemented with proteinase inhibitor (prepared as one proteinase inhibitor pill in 10 ml PBS buffer). Protein extracts were prepared by sonication and subsequent centrifugation at 13500 rpm at 4 degrees Celsius for 30 min. 2 microliter of the supernatant was used for protein quantification by using a NanoDrop 2000c spectrophotometer (Thermo Scientific). Supernatant containing 25 microgram of protein was preincubated with 50 microliter of Amplex Red reagent/HRP working solution prepared according to the kit protocol for 30 min at room temperature, protected from light. The fluorescence was measured with a Varioskan Flash Multimode Reader version 3.00.7 at the excitation wavelength of 540 nm and the emission wavelength 560 nm. The fluorescence intensity was normalized by subtracting the background fluorescence of the Amplex Red reagent/HRP working solution at 30 min.
    Delivered_by: Bacterial_feeding
  Inhibits: Predicted_gene: F46E10.10 Inferred_automatically: RNAi_primary
    Gene: WBGene00018491 Inferred_automatically: RNAi_primary
    Transcript: F46E10.10.1 Inferred_automatically: RNAi_primary
  Species: Caenorhabditis elegans
  Reference: WBPaper00041478
  Phenotype: WBPhenotype:0002350 Remark: Elimination of mdh-1 with RNAi increased the levels of hydrogen peroxide in AMPK mutant dauers
  Remark: (Figure S6B) mdh-1 RNAi. Exact sequence used for RNAi not stated by authors, Ahringer laboratory clone used for curation. If Ahringer clone not available, Vidal laboratory clone used for curation.
  Method: RNAi