RNeasy Lipid Tissue Mini Kit Fragmentation was followed by the synthesis of the first strand cDNA using random hexamer primers, and the second strand cDNA was synthesized using dUTP to ensure strand specificity. The resulting cDNA libraries underwent end repair, A-tailing, adapter ligation, size selection, amplification, and purification. Library quality was confirmed using Qubit for quantification, real-time PCR, and bioanalyzer for size distribution.