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- W2011184654 abstract "Molecular diagnostics based on DNA ampliWcation literally revolutionized biotechnology, health care, forensics, and biodefense [1,2]. Recently emerged techniques that quantify the ampliWed DNA in real time have further advanced these Welds [3–5]. A key element in the real-time ampliWcation methods are Xuorogenic probes, with molecular beacons being among the most useful [4]. Despite an assortment of detection formats, highly potent real-time monitoring of exponential DNA ampliWcation at a single temperature using molecular beacons has not yet been developed. We present here an innovative approach to this important goal. The approach is based on the rolling-circle DNA ampliWcation (RCA), an increasingly popular method for molecular diagnostics due to its simplicity and high speciWcity in the detection of nucleic acids, proteins, and other biomarkers [6]. In addition to DNA polymerase, DNA minicircle and a pair of primers required for exponentially branched RCA [7], our design employs a peptide nucleic acid (PNA) stemless molecular beacon [8,9] and a restriction enzyme, as shown in Fig. 1. Branched RCA reaction yields long double-stranded DNA amplicons generally inaccessible for probe hybridization. Given the unique duplex-invading ability of PNA oligomers [10,11], we assumed that a PNA beacon could rapidly bind the RCA products by strand displacement provided that these products are converted into short" @default.
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- W2011184654 date "2004-12-01" @default.
- W2011184654 modified "2023-10-18" @default.
- W2011184654 title "Real-time monitoring of branched rolling-circle DNA amplification with peptide nucleic acid beacon" @default.
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- W2011184654 doi "https://doi.org/10.1016/j.ab.2004.07.022" @default.
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