Rna labchip rna 6000 nano kit

Total RNA was extracted from cultured cells and ventricular tissues by using TRIzol (Invitrogen, Monza, Italy) and purified by using an RNA purelink mini kit (Invitrogen, Monza, Italy). The concentration and purity of the RNA solution were determined by using a NanoDrop spectrophotometer (Fisher Scientific, Monza, Italy), whereas its overall quality was analyzed using the Agilent 2100 bioanalyzer with an RNA LabChip (RNA 6000 Nano kit, Agilent, Milan, Italy). cDNA was obtained using the High Capacity cDNA Archive kit (Applied Biosystems, Foster City, CA, USA). mRNA expression levels of the β₂-AR were performed using TaqMan gene expression assays (code n. Mm02524224_s1 and Hs00240532_s1 for HL-1 cells and lymphocytes, respectively) (Applied Biosystems, Foster City, CA, USA). qRT-PCR analysis was performed using the 7500 Real-Time PCR system (Applied Biosystems, Foster City, CA, USA). The glyceraldehyde 3-phosphate dehydrogenase (GAPDH, code n Mm99999915_g1 and Hs02786624_g1 for HL-1 cells and lymphocytes, respectively) gene was used as a reference gene, and the ΔCt was used for statistical analysis.

Total RNA was extracted from human cells using TRIzol Plus RNA Purification Kit (AMBION by Life, Technologies, Carlsband, CA, USA). The concentration and purity of the RNA solution was analyzed using the Agilent 2100 bioanalyzer with an RNA LabChip (RNA 6000 Nano kit, Agilent, Milan, Italy). Total RNA was retrotranscribed using the High-Capacity RNA-to-cDNA Kit (Applied Biosystems Carlsbad, CA, USA). Real-time PCR reactions were carried out in 96-well plates using cDNA and TaqMan gene expression assays (Applied Biosystems, Carlsbad, CA, USA) which include specific primers and fluorescent probes for the following genes: β1-ARS (ADRB1, Hs 02330048_s1), β2-ARS (ADRB2, Hs00240532_s1), NOS2 (Hs01075529_m1), PPARɣ (Hs01115513_m1) and GAPDH (Hs 99999905_m1). Real-time PCR was performed on ABI Prism 7500 Fast Sequence Detector (Applied Biosystems).