title: GSM7999305: WT MEF, Normal media, rep1; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA extraction was performed using Trizol (Invitrogen) The illumina RNA libraries for RNA-seq were prepared by TruSeq Stranded mRNA LT Sample Prep Kit according to the manufacturer's protocol. The DNBseq RNA library construction method and sequencing process are carried out according to the following steps: 1. Take a certain amount of total RNA samples, and use oligo dT beads to enrich mRNA with poly A tail; 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.