Not available outside of the UK & Ireland.

Application

Fluorescein-labeled RNA has been used in a variety of hybridization techniques such as: in situ hybridization to tissuesSouthern blotsnorthern blotsplaque or colony liftsRNase protection experiments

General description

Fluorescein RNA Labeling Mix is a convenient nucleotide mixture for the labeling of RNA with fluorescein-12-uridine triphosphate (UTP).

Other Notes

For life science research only. Not for use in diagnostic procedures.

Preparation Note

10x concentrated NTP labeling mixture: 10mM ATP, 10mM CTP, 10mM GTP, 6.5mM UTP, 3.5mM Fluorescein-12-UTP, pH 7.5 (+20°C).

Principle

Fluorescein-labeled, single-stranded RNA probes of defined length are generated by in vitro transcription. Fluorescein-12-UTP is incorporated by SP6, T7, and T3 RNA polymerases at approximately every 20 to 25th nucleotide of the transcript under standard conditions. The Fluorescein RNA Labeling Mix is specifically designed for use with the RNA polymerases SP6, T7, and T3, which are supplied with an optimized transcription buffer.

Quality

The mix is function tested in combination with the DIG RNA Labeling Kit.

Specificity

Heat inactivation: Stop the reaction by adding 2µl 0.2M EDTA (pH 8.0).