HighYield T7 AF647 RNA Labeling Kit, Preparation of randomly AF647-modified RNA probes by in vitro transcription with UTP-PEG5-AF647

Jena Bioscience

HighYield T7 AF647 RNA Labeling Kit, Preparation of randomly AF647-modified RNA probes by in vitro transcription with UTP-PEG5-AF647

5

Research Use Only

HighYield T7 AF647 RNA Labeling Kit is designed to produce randomly AF647-modified RNA probes via in vitro transcription. Such probes are ideally suited for in situ hybridization and Northern Blot experiments. AF647 (a structural analog to Alexa Fluor® 647) is a hydrophilic dye with increased photostability compared to Cy5. UTP-PEG5-AF647 is efficiently incorporated into RNA as substitute for its natural counterpart UTP using an optimized reaction buffer and T7 RNA Labeling Polymerase Mix. 35 % UTP-PEG5-AF647 substitution typically results in an optimal balance between reaction and labeling efficiency. Individual optimization of UTP-PEG5-AF647/UTP ratio however, can easily be achieved with the single nucleotide format. The resulting AF647-modified RNA probe can subsequently be detected by fluorescence spectroscopy. The kit contains sufficient reagents for 20 labeling reactions of 20 microl each (35 % UTP-PEG5-AF647 substitution, 2.5 mM ATP, GTP, CTP, 0.2 mM UTP, 0.1 mM UTP-PEG5-AF647). A 20 microl reaction yields about 4-6 microg RNA after 30 min incubation (1 microg T7 control template, 1.4 kb RNA transcript). Yields may however vary depending on the template (promotor design, sequence length, secondary structure formation).

-20°C

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