GSM9110186: NC_3; Rattus norvegicus; RNA-Seq
Total RNA was extracted using TRIzol Reagent and then treated with RQ1 DNase to remove DNA, followed by assessing its quality and quantity using a smartspec plus spectrophotometer (BioRad) and confirming integrity through 1.5% agarose gel electrophoresis. Each sample underwent mRNA capture using VAHTS mRNA capture Beads (Vazyme, N401), with an additional round of RQ1 DNase treatment. Directional library preparation with the VAHTS Universal V8 RNA-seq Library Prep Kit for Illumina (NR605) involved purifying, fragmenting, and converting polyadenylated mRNAs into double-stranded cDNA. This was followed by end repair, A tailing, and adaptor ligation (N323), with subsequent purification and size fractionation (300 - 500bps). Ligated products were then amplified, purified, quantified, and stored at -80℃ prior to sequencing, with strand-specific sequencing achieved by excluding amplification of the dUTP-marked strand. High-throughput sequencing was performed on an Illumina Novaseq 6000 system using 150nt paired-end sequencing, as per the manufacturer's instructions.