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Real-Time Monitoring in Vitro Transcription Using Molecular Beacons

✍ Scribed by Jianwei Liu; Patricia Feldman; Thomas D.Y. Chung

Publisher
Elsevier Science
Year
2002
Tongue
English
Weight
108 KB
Volume
300
Category
Article
ISSN
0003-2697

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✦ Synopsis

A homogeneous fluorescence-based molecular beacon (MB) method has been developed for real-time monitoring of in vitro transcription reactions. MB probes are structured as target-specific antisense oligodeoxynucleotides containing a proximate fluorophore-quencher pair. Upon binding to its target sequence, the probe undergoes a structural rearrangement that separates the proximate pair, thus dequenching fluorescence. We demonstrate that this simple, inexpensive, rapid, and homogeneous fluorescence-based assay permits real-time monitoring of in vitro transcription and end-point measurement of RNA. The results from the RNA MB assay were comparable to those from other methods.

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