Nanodrop nd 2000 uv vis spectrometer

Total RNA, including miRNAs, was extracted using the miRNeasy minikit (Qiagen, Valencia, CA, USA), following the manufacturer’s instruction. Whole tissues were homogenized before RNA extraction. RNA was quantified with a NanoDrop ND-2000 UV-Vis spectrometer (Thermo Scientific, Wilmington, DE, USA). For rat whole genome miRNA analyses, complementary DNA (cDNA) was synthesized using the Universal cDNA synthesis kit II (Exiqon, Vedbaek, Denmark). miRNAs were analyzed by real time qPCR (qRT-PCR) using the rat miRNome miRNA ready-to-use PCR panels and ExiLENT Sybr green master mix (Exiqon, Vedbaek, Denmark), following the manufacturer’s instructions. miRNome analysis was assessed in 4 samples per group. For individual miRNA analysis, cDNA was synthesized using miScript® II Reverse Transcription kit (Qiagen, Germantown, MD) according to the manufacturer’s guidelines. Individual miRNAs were quantified by qRT-PCR using ABI Prism 7900 Sequence Detection System (Life Technologies, USA). RNU43, RNU6 and/or RNU5G were used as housekeeping genes and the analysis of relative gene expression was analyzed using the 2(-Delta Delta C(T)) method [26 (link)] and using the Exiqon GenEx qPCR analysis software (Exiqon, Vedbaek, Denmark).

DNA was extracted from the target strains using the GenElute™ kit (Sigma, UK), according to the manufacturer’s instructions. The DNA concentration was confirmed using a NanoDrop® ND-2000 UV–vis spectrometer (Thermo Scientific, UK), and the DNA was stored at −20 °C for 6 months.