HighYield T7 Azide RNA Labeling Kit, Preparation of randomly Azide-modified RNA probes by in vitro transcription with 5-Azido-C3-UTP

Jena Bioscience

HighYield T7 Azide RNA Labeling Kit, Preparation of randomly Azide-modified RNA probes by in vitro transcription with 5-Azido-C3-UTP

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Research Use Only

HighYield T7 Azide RNA Labeling Kit is designed to produce 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-C3-UTP is efficiently incorporated into RNA as substitute for its natural counterpart UTP using an optimized reaction buffer and T7 RNA Labeling Polymerase Mix. 35 % 5-Azido-C3-UTP substitution typically results in an optimal balance between reaction and labeling efficiency. Individual optimization of 5-Azido-C3-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[1]. The kit contains sufficient reagents for 25 labeling reactions of 20 microl each (35 % 5-Azido-C3-UTP substitution, 1 mM ATP, GTP, CTP, 0.65 mM UTP, 0.35 mM 5-Azido-C3-UTP).

-20°C

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