GSM8389000: HeLa cells, Control-1; Homo sapiens; RNA-Seq
RNAs were extracted using Trizol. mRNA isolation from total RNA was executed utilizing poly-T oligo-attached magnetic beads. Subsequently, mRNA underwent fragmentation in a buffer containing divalent cations at an elevated temperature. The initial strand of cDNA was synthesized using random hexamers, succeeded by the synthesis of the second cDNA strand. The resultant double-stranded cDNA underwent repair and had an 'A' added to its 3' end. Purification and fragment selection were achieved through Hieff NGS® DNA Selection Beads. Following PCR amplification and enrichment of the products, the target region library underwent denaturation, cycling, and digestion, yielding single-stranded circular DNA. Rolling circle amplification was then employed to amplify the single-stranded circular DNA. The RNA library was sequenced using the MGI T7 platform (BGI Inc., Shenzhen, China), yielding 2 × 150 bp paired-end read datasets.