Cationic conjugated polymers (CCPs) have been widely utilized as signal amplifiers in biosensors to improve the detection sensitivity through fluorescence resonance energy transfer (FRET) from CCPs to dye-labeled probes or targets. This paper investigates the effect of sodium dodecyl sulfate (SDS) on energy transfer between a cationic polyfluoreneethynylene copolymer (P1) and Texas Red labeled single-stranded DNA (ssDNA-TR). The presence of SDS in solution affects both the optical properties of P1 and TR emission within P1/ssDNA-TR complexes, which provides basic information on the role of SDS in FRET between P1 and ssDNA-TR. Although the quantum yield of P1 decreases in the presence of low concentrations of SDS, the presence of SDS reduces TR fluorescence quenching within P1/ssDNA-TR complexes and increases the number of optically active polymer repeat units within the proximity of TR, which are beneficial to P1-sensitized TR emission. In the absence of SDS, FRET from P1 to ssDNA-TR provides a 2.6-fold enhancement in TR emission intensity as compared to that upon direct excitation of TR at 595 nm. At the optimum SDS concentration (5 μM), P1-sensitized TR signal output increases to 11.3-fold relative to direct excitation of TR. This study highlights the importance of modulation of the CCP/ssDNA-dye interaction in improving the signal output of dye-labeled DNA by CCP through FRET