GSM9098563: Tumor tissues, Free PS473 3; Mus musculus; RNA-Seq
RNA was harvested using Rneasy mini plus kit (Qiagen). 1.3 ug of total RNA was usedfor the construction of sequencing libraries. After the RNA samples pass the quality inspection, magnetic beads coated with Oligo(dT) are used to enrich the mRNA of eukaryotes. Subsequently, fragmentation buffer is added to break the mRNA into short fragments. Using the mRNA as a template, first-strand cDNA is synthesized with random hexamers. Then, buffer, dNTPs, DNA polymerase I, and RNase H are added to synthesize second-strand cDNA. After that, AMPure XP beads are used to purify the double-stranded cDNA. The purified double-stranded cDNA undergoes end repair, A-tailing, and sequencing adapter ligation. Then, AMPure XP beads are used for fragment size selection. Finally, PCR amplification is performed, and the PCR products are purified with AMPure XP beads to obtain the final sequencing library.