RNAi was performed as described elsewhere (Fraser et al. 2000 (link); Kamath et al. 2000 ) with minor adaptations when the rrf-3 strain was used: after transferring L3- to L4-staged hermaphrodites onto the first plate, we left them for 48 h at 15°C instead of 72 h and then plated single adults onto other plates seeded with the same bacteria. Furthermore, we did not remove the mothers from the second plates. The phenotypes assayed are these: Emb (embryonic lethal), Ste (sterile), Stp (sterile progeny), Brd (low broodsize), Gro (slow postembryonic growth), Lva (larval arrest), Lvl (larval lethality), Adl (adult lethal), Bli (blistering of cuticle), Bmd (body morphological defects), Clr (clear), Dpy (dumpy), Egl (egg-laying defective), Lon (long), Mlt (molt defects), Muv (multivulva), Prz (paralyzed), Pvl (protruding vulva), Rol (roller), Rup (ruptured), Sck (sick), Unc (uncoordinated) Thin and Pale. Emb was defined as greater than 10% dead embryos for N2 and greater than 30% dead embryos for rrf-3. Ste required a brood size of fewer than ten among fed N2 worms and fewer than five among rrf-3. Each postembryonic phenotype was required to be present among at least 10% of the analysed worms.
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