GSM7866663: MS645_2; Homo sapiens; RNA-Seq
Cells were lysed in Trizol reagent (GE healthcare), and total RNA were isolated according to standard protocal.Three biological replicates were prepared for each treatment. Briefly,  mRNA is enriched by oligo (dT) -attached magnetic beads and fragmented. Reaction systems are prepared to synthesize the double-stranded cDNA. Double-stranded cDNA fragments are subjected to end-repair,  a single 'A' nucleotide attachment and adaptor ligation. The PCR reaction system and program are configured and set up to amplify the product. Single-stranded PCR products are produced via denaturation. The reaction system and program for circularization are subsequently configured and set up. Single-stranded cyclized products are produced, while uncyclized linear DNA molecules are digested. Single-stranded circle DNA molecules are replicated via rolling cycle amplification, and a DNA nanoball (DNB) which contain multiple copies of DNA is generated.