GSM9684294: K181A_3; Homo sapiens; RNA-Seq
For cell line RNA extraction in PromethION full-length transcriptome library prep: lyse cells in TRIzol, add chloroform, centrifuge, and collect the aqueous phase. Precipitate RNA with isopropanol, wash with ethanol, and dissolve in RNase-free water. Use DNase I treatment to remove genomic DNA, then quantify RNA integrity and concentration. The PromethION full-length transcriptome library preparation involves: reverse transcription of poly(A) RNA using a primer with a strand-switching adapter. The resulting cDNA is amplified by PCR, then subjected to adapter ligation (including sequencing adapters and motor proteins). Size selection and quality control steps remove smaller fragments. The final library is loaded onto a PromethION flow cell for long-read, direct RNA or cDNA sequencing, enabling isoform-level transcript analysis.