GSM9278505: MDA-MB-231 cells, si-ATP6V0D2-3; Homo sapiens; RNA-Seq
Total RNA was isolated from three sets of control MDA-MB-231 cells and those treated with siRNA against ATP6V0D2  by TRIzol reagent (Takara, Japan). 1.3ug of total RNA was used for the construction of sequencing libraries. (1) Total RNA samples were tested and then enriched for eukaryotic mRNA by Oligo(dT) magnetic beads; (2) Add Fragmentation Buffer for mRNA fragmentation; (3) One strand of cDNA was synthesised using a six-base random primer using mRNA as a template, followed by the synthesis of the second strand of cDNA using buffer, dNTPs, DNA polymerase I and RNase H, and cDNA purification; (4) Purified double-stranded cDNA was subjected to end repair, base A was added, sequencing junctions were added, and fragments were screened to recover cDNA around 350 bp; (5) PCR enrichment was performed to obtain cDNA libraries.