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Chunk #41 — Materials and methods — Quantitative comparison of RNAi by feeding versus injection

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Effectiveness of specific RNA-mediated interference through ingested double-stranded RNA in Caenorhabditis elegans.
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RNAi was performed by feeding at 15°C using the optimized method described above or by injection. Embryonic lethality was scored by estimating the percentage of dead embryos to the nearest 10% among the offspring of the three worms replica plated for each gene. For embryonic lethality, feeding and injection were considered to be of equal strength if the percentages of dead embryos were within 10%, and one method was considered stronger than the other if the percentage of dead embryos was at least 20% greater than that obtained by the other method. Fed or injected worms were considered sterile if they had fewer than 10 progeny on average, as wild-type worms under similar conditions typically have more than 50 progeny. Post-embryonic phenotypes were scored if more than 10% of the progeny had a given phenotype. For sterility and post-embryonic phenotypes, feeding and injection were considered to be of equal strength if both resulted in that phenotype, and one method was considered stronger if it resulted in that phenotype but the other method did not.