title: GSM8125758: Ctrl_2; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted by Trizol method. A certain amount of RNA was used for the construction of sequencing libraries. The library preparation process involved sample quality control, mRNA isolation and fragmentation, followed by the synthesis of double-stranded cDNA fragments. The cDNA fragments were subjected to end-repair, and then a single 'A' nucleotide was added to the 3' ends of the blunt fragments, followed by adapter ligation reactions. The resulting libraries were PCR-enriched and proceeded for quality control protocol. The subsequent steps included denaturation and circularization to produce single-stranded cyclized products. Single-stranded circle DNA molecules were replicated via rolling cycle amplification to create DNA nanoballs (DNBs), each containing multiple copies of a single molecular entity. Sufficient quality DNBs were then loaded into patterned nanoarrays using high-intensity DNA nanochip technique and sequenced using the combinatorial Probe-Anchor Synthesis (cPAS) method.