GSM1527655: RR_8_2; Diabrotica virgifera virgifera; RNA-Seq
Complete intestinal tracts were removed using forceps, soaked in 50 uL of RNAlater (Ambion, Inc.) and stored (within the solution) at -80°C until use. Total RNA was extracted from these samples using the E.Z.N.A. Total RNA Kit I (Omega Bio-Tek; including DNase treatments). Rootworm populations were grouped into three phenotypic groups based on the documented range where RR-WCR have been reported and previous phenotypic characterization on these populations. The Higginsville (Missouri) and Concord (Nebraska) populations were grouped as WT populations, while the Urbana, Minonk and Shabbona populations from Illinois were classified as RR populations. An additional wild-type population from Ames, Iowa that exhibited a slightly elevated tolerance of soybean diet (yet lower than RR populations) in a previous study was sequenced separately. For each dietary treatment, RNA samples from populations within each “phenotype” were pooled (at equal mass; resulting a "sequencing sample"). For example, one RNA sample from Higginsville WCR fed with corn diets was pooled with another from Concord WCR fed with the same corn diet, resulting an independent “pooled WT x corn” sample. Two biological replicates of such sequencing samples from nine “phenotype x diet” treatments (eighteen in total) were included in this study. RNA-seq libraries were prepared with Illumina's TruSeq RNA Sample Prep kit. Both single and paired-end sequencing were used in this study. For single-end reads, the libraries were quantitated by qPCR, and sequenced on two lanes for 101 cycles on an Illumina HiSeq2000 (Illumina, Inc.) using V3 SBS sequencing chemistry. For paired-end reads, the same libraries were sequenced on one lane for 101 cycles from each end.
GEO Accession
GSM1527655