Real-time monitoring in vitro transcription using molecular beacons

被引:22
作者
Liu, JW [1 ]
Feldman, P [1 ]
Chung, TDY [1 ]
机构
[1] Dupont Merck Pharmaceut Co, Wilmington, DE 19880 USA
关键词
molecular beacon; RNA detection; RNA polymerase; in vitro transcription;
D O I
10.1006/abio.2001.5446
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A homogeneous fluorescence-based molecular beacon (MB) method has been developed for real-time monitoring of in vitro transcription reactions. ME 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 ME assay were comparable to those from other methods. (C) 2001 Elsevier Science.
引用
收藏
页码:40 / 45
页数:6
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