GSM8842538: K7M2col3D_rep1; Mus musculus; RNA-Seq
Total RNA was isolated from the cells by using RNeasy Mini Kit (Qiagen, #74106). 1. Take a certain amount of total RNA samples, and use oligo dT beads to enrich mRNA with poly A tail for DNBSEQ Eukaryotic Strand-specific mRNA library; 2. mRNA molecules were fragmented into small pieces; 3. The fragmented mRNA was synthesized into first strand cDNA using random primers; 4. The second strand cDNA was synthesized with dUTP instead of dTTP; 5. The synthesized cDNA was subjected to end-repair and 3' adenylated. Adaptors were ligated to the ends of these 3' adenylated cDNA fragments; 6. Digest the U-labeled second-strand template with Uracil-DNA-Glycosylase (UDG) and perform PCR amplification; 7. Library quality control; 8. Library circularization; 9. The library was amplified to make DNA nanoball (DNB); 10. Sequencing on DNBSEQ (DNBSEQ Technology) platform.