Journal
BIOSENSORS & BIOELECTRONICS
Volume 77, Issue -, Pages 40-45Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2015.09.027
Keywords
Oligonucleotide; Fluorescence; Target recycling; Exonuclease III; SYBR green I
Categories
Funding
- National Natural Science Foundation of China [21273174]
- Municipal Natural Science Foundation of Chongqing [CSTC-2013jjB00002]
Ask authors/readers for more resources
We describe here a fluorometric assay for sensitive detection of oligonucleotides, based on a target recycling amplification strategy driven by toehold-mediated strand displacement reaction and on exonuclease III (Exo III)-assisted fluorescence background suppression strategy. The network consists of a pair of partially complementary DNA hairpins (HP1 and HP2) with 3' overhang ends, between which the spontaneous hybridization is kinetically hindered by the stems. The target DNA is repeatedly used to trigger a recycling progress between the hairpins, generating numerous HP1-HP2 duplex complexes. Exo III was then employed to digest the double strand parts of the residual hairpins and the intermediate products. The fluorescent dye, SYBR Green I, binds to the double-strand DNA products and emits strong fluorescence to achieve sensitive detection of the target DNA with the detection limit of 5.34 pM. Moreover, this proposed strategy showed high discrimination efficiency towards target DNA against mismatched DNA and was successfully applied in the analysis of human serum sample. (C) 2015 Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available