title: RNA-seq
design: Total RNA was processed using mRNA enrichment method. mRNA enrichment: PolyA tail mRNA was enriched using magnetic beads with OligodT. Use interrupt buffer to fragment the obtained RNA, perform reverse transcription with random N6 primers, and then synthesize cDNA double stranded DNA. Flatten the ends of the synthesized double stranded DNA and phosphorylate the 5 'end to form a sticky end with a protruding "A" at the 3' end, and then connect it to a bubbly junction with a protruding "T" at the 3 'end. The connecting product is amplified by PCR using specific primers. The PCR product undergoes thermal denaturation to form a single stranded DNA, which is then cyclized using a bridge primer to obtain a single stranded circular DNA library. The constructed library undergoes quality inspection and sequencing after passing the inspection.
identifiers.namespace: SUB15930920
identifiers.id: 1