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Anal Biochem. 2002 Jan 1;300(1):40-5.

Real-time monitoring in vitro transcription using molecular beacons.

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  • 1DuPont Pharmaceuticals, Wilmington, Deleware 19880, USA. jianwei.liu@astrazeneca.com


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.

(c)2001 Elsevier Science.

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