title: GSM8630823: youth-2, 6M; Mus musculus; RNA-Seq
library_construction_protocol: RNA was harvested using traditional TRIzol methods. After sample quality inspection, take a certain amount of RNA sample, denature at appropriate temperature to open its secondary structure, and use oligo (DT) magnetic beads to enrich mRNA. Add interrupting reagent to the mRNA obtained in the previous step, react at an appropriate temperature for a certain time, and fragment the mRNA. Prepare one strand synthesis reaction system and set the reaction procedure to synthesize one strand cDNA; The second strand synthesis reaction system was prepared, and the reaction procedure was set to synthesize the second strand cDNA. The reaction system was prepared by end repair, adding "a" and linker connection, and the reaction procedure was set to repair the ends of double stranded cDNA, and adding a base at the 3 'end; The linker ligation reaction system was prepared, and the reaction procedure was set to connect the linker with cDNA. The PCR reaction system was prepared, and the reaction procedure was set to amplify the product. After the quality inspection of the library, the PCR product was denatured into a single strand, the cyclization reaction system was prepared, and the reaction procedure was set to obtain a single stranded circular product, which digested the linear DNA molecules that were not cyclized. Single stranded circular DNA molecules replicate through rolling rings to form a DNA nanosphere (DNB) containing multiple copies. The obtained DNBS were added into the mesh pores on the chip using high-density DNA nano chip technology, and sequenced by combined probe anchor polymerization (CPAs).