RNA-Seq of Culex quinquefasciatus: UT
Initially, DNA was isolated from tissue samples using the DNeasy PowerSoil Pro Kit (Qiagen, Cat #47014). The quantity and quality of the isolated DNA were then assessed using a Nanodrop spectrophotometer (Thermofisher Scientific) and confirmed by gel electrophoresis (Lonza, Cat #50004), ensuring the DNA was suitable for downstream applications. For library preparation, the KAPA HyperPlus Kit (Roche, Cat #KR1145-v8.21) was employed, which involves DNA fragmentation, end repair, adapter ligation, and optional amplification, following standard next-generation sequencing (NGS) protocols to generate sequencing-ready libraries. The KAPA HyperPure beads (Roche, Cat #KK8009) were used for cleanup and size selection during library preparation to improve library quality. Library quantification was performed using the Qubit 1X dsDNA High Sensitivity Kit (Invitrogen, Cat #2491434) and further quality-checked with the Agilent Bioanalyzer High Sensitivity DNA kit (Cat #2215) to confirm fragment size distribution and concentration. Finally, sequencing was carried out on the Illumina NovaSeq 6000 platform using the S4 Flow Cell.