Two New Kits Designed to Promote the Study of RNA-Protein Interactions

Two related self-contained kits allow molecular biologists to probe RNA-protein complexes using a modernized version the RNA electrophoretic mobility shift assay (EMSA) technique that replaces radioactive labeling with chemiluminescent detection.

The kits are manufactured by two subdivisions of Thermo Fisher Scientific Inc. (Milford, MA, USA). The first is the LightShift RNA EMSA Kit, which the manufacturer says is the first commercially available EMSA kit. The protocol is based on a biotinylated RNA fragment used to probe protein samples. RNA-protein complexes are then monitored by changes in electrophoretic migration patterns. To detect these changes, a chemiluminescent detection method is used that provides sensitivity comparable to detection of a radioactive label.

Preparing the biotinylated probe in-house is made easy with the second kit, the Thermo Scientific Pierce RNA 3' End Biotinylation Kit. This kit provides the reagents required to produce biotinylated RNA probes for use in chemiluminescent or other detection methods. An enzymatic procedure is used to attach a single biotinylated nucleotide to the 3′ end of an RNA strand that is 22 to 450 nucleotides in length. The reaction can be completed in 30 minutes to two hours.

The biotinylated RNA probes produced in-house cost much less than commercial biotinylated RNA probes. The complete kit contains an unlabeled RNA strand to use as a positive control, a biotinylated-RNA probe standard, RNase inhibitor, glycogen, and ligation-enhancing reagents.

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