Journal
TALANTA
Volume 115, Issue -, Pages 819-822Publisher
ELSEVIER
DOI: 10.1016/j.talanta.2013.06.062
Keywords
DNA machine; Homogeneous; Fluorescence; Lambda exonuclease; Signal amplification
Categories
Funding
- National Basic Research Program of China [2010CB732400]
- National Natural Science Foundation of China [21075060, 21121091, 21135002]
- Program for New Century Excellent Talents in University [NCET100479]
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This work designs a DNA machine with three assistant DNAs and lambda exonuclease (Exo-lambda) for sensitive and homogeneous fluorescent detection of DNA. The selective digestion of Exo-lambda to blunt or recessed 5'-phosphorylated strand of probe 1-probe 2 duplex results in the release of target DNA and probe 2 to produce the fluorescence restoring of fluorophore labeled to probe 1. The released target DNA could hybridize with another probe 1-probe 2 duplex to trigger the target recycling for signal amplification, while the released probe 2 hybridized with molecular beacon to restore its fluorescence for signal enhancement. This DNA machine showed a fast response to target DNA with a linear concentration range from 0.4 pM to 4 nM. The limit of detection was 68 fM at a signal-to-noise ratio of 3. The high selectivity of the method may result from the Exo-lambda's specific recognition-site of double-stranded DNA and the specific hybridization of target DNA with probe 1-probe 2 duplex. This DNA machine with the homogenous detection, rapid response as well as simplicity provides a new approach for sensitive detection of DNA. (C) 2013 Elsevier B.V. All rights reserved.
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