HighYield T7 Azide RNA Labeling Kit (UTP-based)

HighYield T7 Azide RNA Labeling Kit (UTP-based) is designed to pro-duce randomly Azide-modified RNA probes via in vitro transcription. Such probes are ideally suited for in situ hybridization, Northern Blot or crosslinking experiments. 5-Azido-C 3-UTP is efficiently incorporated into RNA as substi-tute for its natural counterpart UTP using an optimized reaction buffer and T7 RNA Labeling Polymerase Mix. 35% 5-Azido-C 3-UTP substitution typically results in an optimal balance between reaction and labeling efficiency. Individual optimization of 5-Azido-C 3-UTP/UTP ratio however, can easily be achieved with the single nucleotide format. The resulting Azide-modified RNA probe can subsequently be labeled by copper-free Click Chemistry with Dibenzocyclooctyne (DBCO) -functionalized detection reagents (e.g DBCO-functionalized fluorescent dyes or (Desthio) Biotin) thereby forming a stable, covalently labeled RNA probe. The kit contains sufficient reagents for 25 labeling reactions of 20 μl each (35% 5-Azido-C 3-UTP substitution, 1 mM ATP, GTP, CTP, 0.65 mM UTP, 0.35 mM 5-Azido-C 3-UTP) .