RGB colour coding of Y-shaped DNA for simultaneous tri-analyte solid phase hybridization detection.
We present a new concept for tri-analyte DNA detection based on the idea of a Y-shaped capture probe which, after tri-target and fluorescently labeled reporter probe binding, becomes colour-coded to generate images in an RGB colour scheme. Hence, the RGB value of the resulting secondary pseudo-colour presented by the hybridized Y-DNA can be related to the ratio of the primary pseudo-colours present in its make-up, and thus to the ratio of the three target concentrations. As a proof of concept we detect sequences from the genes of the pathogenic bacterial strains Escherichia coli O157:H7, Vibrio cholera and Salmonella enteric in a semi-quantitative manner across the range 20-167 nM. The assay was relatively quick, with a time from hybridization to completed data interpretation of approximately 4 h.
School of Bioresources and Technology, King Mongkut's University of Technology Thonburi, Bangkhuntien Campus, Bangkok 10150, Thailand.
Abhichart
A
Krissanaprasit
Mithran
M
Somasundrum
Werasak
W
Surareungchai
D015374:Biosensing Techniques / Q000295:instrumentation*; D003116:Color; D003124:Colorimetry / Q000295:instrumentation*; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation*; D044367:Phase Transition
D015342:DNA Probes; D004269:DNA, Bacterial