Incorporation of metalated nucleosides into DNA through covalent modification is crucial to measurement of thermal electron-transfer rates and the dependence of these rates with structure, distance, and position. Here, we report the first synthesis of an electron donor-acceptor pair of 5' metallonucleosides and their subsequent incorporation into oligonucleoticles using solid-phase DNA synthesis techniques. Large-scale syntheses of metal-containing oligonucleoticles are achieved using 5' modified phosporamidites containing [Ru(acac)(2)(IMpy)](2+) (acac is acetylacetonato; IMPy is 2'-iminomethylpyridyl-2'-deoxyuridine) (3) and [Ru(bpy)(2)(IMpy)](2+) (bpy is 2,2'-bipyridine; IMPy is 2'-iminomethylpyridyl-2'-deoxyuridine) (4). Duplexes formed with the metal-containing oligonucleoticles exhibit thermal stability comparable to the corresponding unmetalated duplexes (T-m of modified duplex = 49 degreesC vs T-m of unmodified duplex = 47 degreesC). Electrochemical (3, E-1/2 = -0.04 V vs NHE; 4, E-1/2 = 1.12 V vs NHE), absorption (3, lambda(max) = 568, 369 nm; 4, lambda(max) = 480 nm), and emission (4, lambda(max) = 720 nm, tau = 55 ns, Phi = 1.2 x 10(-4)) data for the ruthenium-modified nucleosides and oligonucleoticles indicate that incorporation into an oligonucleotide does not perturb the electronic properties of the ruthenium complex or the DNA significantly. In addition, the absence of any change in the emission properties upon metalated duplex formation suggests that the [Ru(bpy)(2)(IMPy)](2+)[Ru-(acac)(2)(IMPy)](2+) pair will provide a valuable probe for DNA-mediated electron-transfer studies.