GSM8138806: ABE8e_rep1_HEK293T; Homo sapiens; RNA-Seq
Total RNA is extracted utilizing the Trizol method Total RNA is extracted utilizing the Trizol method, subsequently assessed for purity using NanoDrop One, and its integrity is evaluated with Agilent 2100.Following this, mRNA enrichment and purification from eukaryotic total RNA are executed with the VAHTS mRNA Capture Beads kit (Vazyme), and RNA is fragmented through ion interruption to attain insert sizes ranging from 250 to 450 bp. cDNA first-strand synthesis is conducted using fragmented RNA as a template, succeeded by second-strand cDNA synthesis employing the first-strand cDNA as a template, and subsequent ds cDNA end repair and dA-tailing. Following the ligation of universal adapters, bead-based purification is utilized, and fragment selection is executed for sizes ranging between 250 and 350 bp. PCR amplification is conducted, incorporating primer double-end indexes, and the products undergo bead-based purification to yield the complete library. Second-generation sequencing technology, leveraging the Illumina NovaSeq 6000 sequencing platform, is employed for paired-end (PE) sequencing of the library.