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Four biological replicate pools were generated for each of the 11 states.
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The number of biological replicate pools was four for each sampling point.
Each biological replicate pool was co-hybridized in a two-dye experiment with a single pooled reference sample.
From each biological replicate, three independent pools were created using 100 ng total RNA from individual insects.
At each time point, three biological replicates were designed; the samples of each biological replicate were pooled from 10 plants, the plants being randomly selected to avoid any potential effects of position within the growth room.
After extraction, total RNAs from each biological replicate were pooled in equal quantity to obtain a total RNA mixture representative of 180 individuals.
Equal amounts of total RNA from each biological replicate were pooled and used for isolation of the small RNA fraction using the miRVana™ miRNA Isolation kit (Ambion®).
Four μg of each biological replicate were pooled together and freeze-dried.
Six libraries from a biological replicate were pooled on the same HiSeq2000 lane.
Three independent preparations of mRNA for each biological replicate were pooled to eliminate inconsistencies due to sampling.
Three independent preparations of mRNA for each biological replicate were pooled to eliminate the inconsistent variation with sampling.
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