GSM8703109: NSC treated with ultrasound, sample2 of C grup, Day5; Rattus; RNA-Seq
The BGI Optimal series dual module mRNA library construction kit (BGI Shenzhen, China) was used for library preparation. Take a certain amount of RNA sample, perform temperature denaturation to open the secondary structure, and use oligo (dT) magnetic beads to enrich mRNA. At an appropriate temperature, add a disruption reagent to the mRNA obtained in the previous step, and perform a temperature appropriate reaction to fragment it.                                                                                                                                                                                        Then, the first strand cDNA was generated by reverse transcription using random hexamer primers, followed by the synthesis of the second strand cDNA. Afterwards, prepare the reaction system, repair the double stranded cDNA end, and add A base at the 3 'end. Prepare a linker connection reaction system, connect the linker to cDNA, and finally amplify and quality control the product.                                                      After denaturing the PCR product into a single strand, a cyclization reaction system is prepared and a reaction program is set up to obtain a single stranded circular product. The linear DNA molecules that have not been cyclized are digested to obtain the final library. The final library was amplified by phi29 to produce DNA nanospheres (DNBs). The obtained DNBs were added to the mesh like pores on the chip using high-density DNA nanochip technology, and PE100/PE150 sequencing was performed on a G400/T7/T10 sequencer (BGI Shenzhen, China) using co probe anchoring polymerization technology (cPAS).