TPE Treated B
Total RNAs were extracted from the TPE treated Culex quinquefasciatus larvae replicate3, using TRIzol reagent and quantified using NanoDrop 2000 (Thermo Inc., Massachusetts, USA). The integrity of the extracted RNA was assessed by gel electrophoresis. To minimize cross-contamination, all RNA and DNA analyses were conducted in separate designated areas. The RNA gel apparatus was used exclusively for RNA analysis, along with dedicated loading dyes and RNA markers. Additionally, nuclease-free TAE buffer was diluted with DEPC-treated water to maintain RNA integrity.cDNA synthesis and library preparation were performed using the KAPA HyperPrep Kit for cDNA Synthesis & Amplification Module (Roche, Switzerland). Synthesized cDNA was purified with KAPA Pure Beads (Roche). Library quality and size distribution were verified on an Agilent Bioanalyzer (Agilent Technologies, USA). Sequencing was carried out on the NovaSeq 6000 platform (Illumina, USA) to generate paired-end reads.