HighYield T7 AF555 RNA Labeling Kit, Preparation of randomly AF555-modified RNA probes by in vitro transcription with UTP-PEG5-AF555

Catalogue Number: RNT-101-AF555-JEN

Manufacturer:Jena Bioscience
Shelf Life:12 months
Type:Nucleic Acid Labeling Kits
Shipping Condition:Blue Ice
Unit(s): 20 reactions

Description

Description: HighYield T7 AF555 RNA Labeling Kit is designed to produce randomly AF555-modified RNA probes via in vitro transcription. Such probes are ideally suited for in situ hybridization and Northern Blot experiments. AF55 (a structural analog of Alexa Fluor® 555) is a hydrophilic dye with increased photostability compared to Cy3. UTP-PEG5-AF555 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-AF555 substitution typically results in an optimal balance between reaction and labeling efficiency. Individual optimization of UTP-PEG5-AF555/UTP ratio however, can easily be achieved with the single nucleotide format. The resulting AF555-modified RNA probe can subsequently be detected by fluorescence spectroscopy. The kit contains sufficient reagents for 20 labeling reactions of 20 µl each (35 % UTP-PEG5-AF555 substitution, 2.5 mM ATP, GTP, CTP, 0.2 mM UTP, 0.1 mM UTP-PEG5-AF555). A 20 µl reaction yields about 5-7 µg RNA after 30 min incubation (1 µg T7 control template, 1.4 kb RNA transcript). Yields may however vary depending on the template (promotor design, sequence length, secondary structure formation).

Additional Text

Smile String

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