RNA-Seq of Caenorhabditis elegans: Treatment
RNA was denatured to open its secondary structure, mRNA was enriched with oligo(dT) magnetic beads, and cDNA was synthesized after fragmentation. End repair, add "A" and joint connection; PCR reaction system was prepared to amplify the product. Quality inspection of the library; After the PCR product was denatured to single strand, the cyclization reaction system was prepared to obtain the single strand ring product, and the linear DNA molecules that were not cyclized were digested. Single-stranded circular DNA molecules are replicated by rolling rings to form a DNA nanosphere (DNB) containing multiple copies. The DNBs obtained were added to the mesh holes on the chip using high-density DNA nanochip technology, and sequenced by combined probe anchoring polymerization (cPAS).