Comparison of RNA extraction by Arpengen kit with RNA extraction by TRIzol and the subsequent use of the kit in a gene expression study

Abstract

RNA used in gene expression studies must be free of contaminants and sufficiently intact to obtain reliable results. This work compares the Arpengen kit with a membrane column and the Trizol solution, on tissues that are difficult to process estimating the RNA quality using spectrophotometry, gel electrophoresis and RT-qPCR. After determining that the kit isolates RNA with sufficient quality, is user-friendlier and the result more reproducible than TRIzol extraction, it was used to isolate RNA from the prefrontal cortex of mice for gene expression analysis to investigate whether the Negr1 gene interacts with selected components of the endocannabinoid system and the blood-brain barrier molecule claudin-5. The results showed that Negr1-deficiency affects the expression level of cannabinoid receptors and claudin-5 in response to acute high-fat diet exposure.