Silkworm Antennal Transcriptome Analysis

Total RNA was extracted from 10 silkworm antennae from 5 males per genotype using Trizol reagent (Invitrogen) and treated with RNase-free DNAse I (Ambion). cDNAs were synthesized using the Omniscript Reverse transcriptase kit (Qiagen) in a 20 μl reaction mixture containing 1 μg total RNA. Quantitative real-time RT-PCR (RT-qPCR) assays were performed using SYBR Green Realtime PCR Master Mix (Thermo Fisher Scientific) on an Eppendorf Real-time PCR System MasterCycler RealPlex instrument. RT-qPCR reactions were carried out with gene-specific primers (S1 Table). A 10-fold serial dilution of pooled cDNA was used as the template for standard curves. Quantitative mRNA measurements were performed in three independent biological replicates, and data were normalized to the amount of Bmrp49 mRNA [12 (link)].