Labelit mirna labelling kit

MicroRNA profiling has been described previously in detail [3 (link),4 (link),5 (link)], which we reproduce in quotes as follows: “In brief, total RNA and miRNAs were extracted using the Trizol standard protocol (Invitrogen, Carlsbad, CA, USA) and the mirVANA miRNA isolation kit (Ambion, Austin, TX, USA). Labelling and hybridization were performed using the LabelIT miRNA labelling kit (Mirus Bio LLC, Madison, WI, USA) according to manufacturer’s instructions. Samples were hybridized to Applied MicroArrays (miRlink Bioarray 300054-3PK) platform. This array contained 1211 human miRNAs. Hybridization was performed at 37 °C with rotation at 145 rpm for 16 h. Images were scanned using Agilent Microarray Scanner (G2565CA) controlled by Agilent Scan Control 7.0 software. The total gene signals were extracted using the Imagene 6.0 software (Biodiscovery Inc., El Segundo, CA, USA) that contains summarized signal intensities for each miRNA by combining intensities of replicate probes and background subtraction” [3 (link),4 (link),5 (link)]. In total, 49 CNS tumor samples (the complete cohort described in Section 2.1.) and 13 control samples were investigated for their miRNA expressional profile. All microarray data are MIAME compliant.

In brief, total RNA and miRNAs were extracted using the Trizol standard protocol (Invitrogen, Carlsbad, CA) and the mirVANA miRNA isolation kit (Ambion, Austin, TX). The RNA quantity and quality were evaluated using a spectrophotometer (NanoDrop® ND-1000 UV–vis, Nanogen Inc.). Labelling and hybridization were performed using the LabelIT miRNA labelling kit (Mirus Bio LLC, USA) according to manufacturer’s instructions. Samples were hybridized to Applied MicroArrays (miRlink Bioarray 300054-3PK) platform. This array contained 1211 human miRNAs. Hybridization was performed at 37°C with rotation at 145 rpm for 16 h. Images were scanned using Agilent Microarray Scanner (G2565CA) controlled by Agilent Scan Control 7.0 software.