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Fig. 6 Example of a positive suppression interaction. (A-B) Repeats of the control for the mutation:
emb-30(g53) at 25 C is lethal. (C-D) Suppression of lethality of emb-30(g53) at 25 CbytheRNAiof
the gene smg-1, repeats A and B, respectively. Insets are enlarged sections of the pictures to show
details.
C. Quantification of Embryonic Lethality or Survival on Agar Plates
The positive results in the large-scale screen in liquid for enhancement or sup-
pression can be repeated on agar plates to confirm and quantify the results.
Day 1
Replicate bacteria from 80 C stock to LB agar with 50 mg/L ampicillin and
10 mg/L tetracycline rectangular plates and place overnight at 37 C in a dry
incubator.
Day 2
Grow bacteria in liquid LB with 50 mg/L ampicillin shaking at 250 rpm for 12-16
h, but no more than 18 h.
Day 3
1. For each worm strain-RNAi clone pair to be tested (including the empty vector,
L4440), seed three RNAi plates with 30 m L of bacteria and one RNAi plate
with 300 m L of bacteria, and let dry for 24 h.
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