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To carry out the experiment based on the separate reduction and derivatization reactions the set of samples were prepared as follow

For gene expression analysis, 1 g of RNA was used for cDNA synthesis using qScript cDNA Synthesis Kit (Quanta Bio 66196756)

Singh SJ, Puhan MA, Andrianopoulos V, Hernandes NA, Mitchell KE, Hill CJ, et al

Key Features & Research Applications Fat Reduction & Lipid Metabolism: Shown to decrease fat cell size and inhibit fat accumulation by blocking NNMT enzyme activity