title: GSM9253765: HCT116 cells, shCtrl-2; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted using the Trizol Reagent (B610409, Sangon Biotech). The total RNA was processed using the mRNA enrichment method. mRNA enrichment: mRNA with polyA tails was enriched using magnetic beads coated with Oligo(dT). The obtained RNA was fragmented using fragmentation buffer, followed by reverse transcription with random N6 primers to synthesize cDNA second strands, forming double-stranded DNA. The ends of the synthesized double-stranded DNA were blunted and phosphorylated at the 5' end, while the 3' end formed a sticky end with an protruding "A" base. A bubble-shaped adapter with a protruding "T" at the 3' end was then ligated. The ligation products were amplified by PCR using specific primers. The PCR products were heat-denatured into single strands, and a bridge primer was used to circularize the single-stranded DNA, resulting in single-stranded circular DNA libraries. The constructed libraries underwent quality inspection, and those passing the quality control were subjected to sequencing.