NDLI: Using molecular beacons as a sensitive fluorescence assay for enzymatic cleavage of single-stranded DNA.

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Using molecular beacons as a sensitive fluorescence assay for enzymatic cleavage of single-stranded DNA.

Content Provider	Semantic Scholar
Author	Li, Jianwei Geyer, Rudolf Tan, Weihong
Copyright Year	2000
Abstract	Traditional methods to assay enzymatic cleavage of DNA are discontinuous and time consuming. In contrast, recently developed fluorescence methods are continuous and convenient. However, no fluorescence method has been developed for single-stranded DNA digestion. Here we introduce a novel method, based on molecular beacons, to assay single-stranded DNA cleavage by single strand-specific nucleases. A molecular beacon, a hairpin-shaped DNA probe labeled with a fluorophore and a quencher, is used as the substrate and enzymatic cleavage leads to fluorescence enhancement in the molecular beacon. This method permits real time detection of DNA cleavage and makes it easy to characterize the activity of DNA nucleases and to study the steady-state cleavage reaction kinetics. The excellent sensitivity, reproducibility and convenience will enable molecular beacons to be widely useful for the study of single-stranded DNA cleaving reactions.
Starting Page	E52
Ending Page	E52
Page Count	1
File Format	PDF HTM / HTML
Alternate Webpage(s)	https://oup.silverchair-cdn.com/oup/backfile/Content_public/Journal/nar/28/11/10.1093_nar_28.11.e52/1/2800e52.pdf?Expires=1492574625&Key-Pair-Id=APKAIUCZBIA4LVPAVW3Q&Signature=ACidi4bJUbT~L6BLXMbFFw7-YYhzN1-K3wfyaSGT5ajzRZpAQYzpPHqEABqYuZtFGU5HlGc08F2hua2XjsZmRzPlXD8Qh8b7TVPcxeQvIQfIUEo66OeL~9E4u6nALLI5rjVrK4QKiboKC43cDDkOdLCrY-SmfCAcJihr14vXhmI1Hy8EFcINLCHojz~D~Uk9O31YdqlqNhGHBEwqCaAvediiK4Jhb5psBu1kSHsBPF~e-3oNGU5dhYcZWoSZ2b1EYn23tK2m9FO271ItfaMeIMmtARXXISaIoTMfkA3djI38ZmIay78azIKd~IQV5A9pu9nEyh4vrs2LqpbOugh09A__
PubMed reference number	10871351v1
Volume Number	28
Issue Number	11
Journal	Nucleic acids research
Language	English
Access Restriction	Open
Subject Keyword	DNA Breaks, Single-Stranded DNA Cleavage DNA Probes Deoxyribonucleases Fluorescence License Nucleic Acid Strand Tracer nuclease
Content Type	Text
Resource Type	Article

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