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Total RNA was extracted from mosquitoes using the RNeasy Mini Kit (Qiagen, Germany), following the manufacturer's protocol. First-strand complementary DNA (cDNA) was synthesized via reverse transcription using random hexamer primers and the Illumina TruSeq Stranded Total RNA Kit (Illumina Inc., USA), ensuring comprehensive coverage of the RNA template. Second-strand synthesis was performed using the same kit to generate double-stranded cDNA (ds-cDNA), providing a stable representation of the transcriptome. The resulting ds-cDNA was used for library construction using the Illumina DNA Prep Kit, and indexed libraries were pooled at equimolar concentrations. The pooled libraries were normalized to 4 nM and diluted to a final loading concentration of 1.5 pM before paired-end sequencing on the Illumina NextSeq 550 platform. The resulting high-throughput sequence data served as the basis for metatranscriptomic profiling. |
RNA-Seq |
TRANSCRIPTOMIC |
cDNA |
PAIRED
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