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The dissected fresh Bb gonads were homogenized and lysed using TRIzol, stored at -80C, and subsequently shipped on dry ice to Novogene for sequencing. Total RNA was extracted from the gonads of Bb type b females and males using TRIzol, following the RNA-seq protocol. The extracted RNA served as the input material for RNA sample preparation. Briefly, 3' and 5' adaptors were ligated to the respective ends of small RNA molecules. First-strand cDNA synthesis was performed after hybridization with a reverse transcription primer. Subsequently, a double-stranded cDNA library was generated through PCR enrichment. Libraries with inserts ranging from 18 to 40 base pairs were purified, size-selected, and prepared for sequencing. After completing library construction, the inserted fragments were quantified to ensure effective concentration and high-quality libraries. The qualified libraries were pooled and sequenced on Illumina platforms. |
miRNA-Seq |
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other |
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