Identification of active target genes in bioassay screening is the first important step for application of RNA interference (RNAi) for pest control. Here, we describe the methodology for performing high-throughput RNAi target screening against important agriculture pest, Western corn rootworm in 96-well microplate. Two approaches are presented to identify active targets from random-cDNA library or testing a certain group of specific targets via in silico sequence analysis. Methods of PCR primer design, DNA template preparation, and dsRNA production described here can be applied for other pests.
Keywords: Diabrotica virgifera virgifera; Diet assay; Double-stranded RNA (dsRNA); In vitro transcription (IVT ); RNA interference (RNAi); Western corn rootworm (WCR ).
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