DNA quantification based on FRET realized by combination with surfactant CPB.
In this work, we developed a novel DNA quantitative analysis based on fluorescence resonance energy transfer (FRET) realized by combination with a surfactant CPB. The approach was capable of detecting long-stranded DNA in a separation-free format. A sandwich-type FAM-c-DNA-t-DNA-r-DNA-TAMRA conjugate was first formed by the capture probe tagged with FAM, the reporter probe tagged with TAMRA and the target DNA through hybridization. The donor (FAM) and the acceptor (TAMRA) were bridged to afford a FRET system. Subsequently, an addition of the cationic surfactant CPB to the system resulted in a substantial change of the microenvironment and an effective condensation of DNA strands. Consequently, without altering the component of the double strands, an enhanced acceptor fluorescence signal from FRET was achieved and a quantification of the target DNA containing 30 bases was enabled. Under the optimal experimental conditions, an excellent linear relationship between the increase of acceptor fluorescent peak area and the target DNA concentration was obtained over the range from 1.0 x 10(-7) to 3.0 x 10(-9) mol L(-1). The proposed approach offered adequate sensitivity for the detection of the target DNA at 1.0 x 10(-9) mol L(-1).
Liu
Chunxia
C
School of Chemistry and Chemical Engineering, Shandong University, 250100 Jinan, PR China.
Wang
Lei
L
Jiang
Wei
W
D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D031541:Fluorescence Resonance Energy Transfer; D057230:Limit of Detection; D009690:Nucleic Acid Conformation / Q000187:drug effects; D009693:Nucleic Acid Hybridization; D013501:Surface-Active Agents / Q000737:chemistry* / Q000494:pharmacology
D015342:DNA Probes; D004269:DNA, Bacterial; D013501:Surface-Active Agents; C036483:T-DNA; D004247:DNA