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Total RNA is extracted from blood samples using TRIzol reagent, followed by an assessment of RNA concentration, purity, and integrity using Nanodrop 2000 and Agilent 2100 bioanalyzer. Subsequently, mRNA is enriched from total RNA using magnetic beads with Oligo(dT), fragmented, and reverse-transcribed into cDNA using random hexamers. Double-stranded cDNA is synthesized using DNA polymerase I and RNase H, purified with AMPure XP beads, and subjected to end repair, A-tailing, adapter ligation, and size selection with AMPure XP beads. |
RNA-Seq |
TRANSCRIPTOMIC |
PCR |
PAIRED
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