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The total RNA sample was first enriched for polyA-tailed mRNA using oligo dT beads, followed by fragmentation of the mRNA into small pieces. The fragmented mRNA was then reverse-transcribed into first-strand cDNA using random primers, followed by second-strand cDNA synthesis. The resulting double-stranded cDNA underwent end repair, 3' adenylation, and adapter ligation. After PCR amplification and purification, the library was subjected to quality control, circularized, and finally amplified to generate DNA nanoballs (DNBs) for sequencing. |
RNA-Seq |
TRANSCRIPTOMIC |
PCR |
PAIRED
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