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The feeding RNAi was performed at 20°C for ~55 hours, during which the feeding plate was exchanged once after the first 24 hours.
We defined feeding time (number of hours of RNAi) as the time between transfer of worms to the feeding plate and putative fertilization of the egg.
We grew the worms at 15°C to minimize genomic instability and we transferred 5 6 larvae of each genotype to a new feeding plate every two generations.
We thus predicted that latency to initially visit a feeding plate would decrease and the number of visits would increase with increasing distance from the observer.
We expected that weaver behavior would vary with this increased perception of risk such that the latency to initially visit a feeding plate would increase with increasing distance from cover, and the number of visits would decrease.
Likewise, narrow birth interval [COR 95% CI) = 1.60 (1.2.092.09)], lack of separate feeding plate for the child [COR 95% CI) = 1.40 (1.89, 1.89)], and feeding a child from less than four major food items within 24 hours preceding the survey [COR 95% CI) = 1.53 (1.17, 2.00)] were identified as individual level factors that are associated with children's nutritional status (Table 2).
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Worms were grown for 4 30 h (according to the individual dsRNA) at 25°C on feeding plates, or on normal plates after injection.
For hda-3, a 500 bp fragment of cDNA was amplified by PCR and cloned in the feeding vector pPD129.36 using following primers: hda3up: 5'-ACGCGTCGACATGAGCCTCCAACACTCGAAATC-3'-CATGCCATGGTGATGCTTCAAAAGCTCCAAAATC-3GCTCCAAAATC-3' L4 worms were put on RNAi feeding plates at 25°C and their progeny was scored for synMuv phenotype.
With approximately 1.1×104 packed L1 animals per µl (n = 1) and typical yields of approximately 40 µl of packed worms from three 100 mm diameter feeding plates seeded with a full lawn of bacteria, we obtained 1.1±0.8×106 cells during a typical L1 isolation, or approximately 2.5±1.7 cells per animal.
RNAi-treated worms were placed on RNAi feeding plates starting at the first day of adulthood.
lin-35 ; slr-2 ; kuEx119 larvaevae were placed on vector-control or suppressor RNAi feeding plates and allowed to develop into adults.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com