GSM8956236: SK-N-SH cells, LINC02449-rs149707223_Ref-C OE, rep2; Homo sapiens; RNA-Seq
RNA was harvested by the phenol-chloroform method using TRIZOL (Invitrogen). 1 ug of total RNA was used for the construction of sequencing libraries. Using magnetic beads coated with Oligo(dT), mRNA with polyA tails is enriched and purified from RNA samples. Fragmentation Buffer is then added to the purified mRNA to fragment it. The resulting RNA fragments serve as templates for the reverse transcription process, utilizing random primers to synthesize the first strand of cDNA. Subsequently, a 2nd Strand Marking Buffer and a 2nd Strand/End Repair Enzyme Mix are introduced to synthesize the second strand of cDNA. Following this, end repair is performed, an adenine base is added, and sequencing adapters are ligated. The target fragments are then recovered through magnetic bead selection and subjected to PCR amplification, thereby completing the entire library preparation process.