In this work, it is shown that the incorporation of an 8-deuteroguanine (G*) moiety in DNA-oligomers allows for direct determination at 77 K of (i) the location of holes (i.e., the radical site) within dsDNA at specific base sites, even within stacks of G, as well as (ii) the protonation state of the hole at that site. These findings are based on our work and demonstrate that selective deuteration at C-8 on a guanine moiety in dGuo results in an ESR signal from the guanine cation radical (G**(+)) which is easily distinguishable from that of the undeuterated guanine cation radical (G*(+)). G**(+) is also found to be easily distinguishable from its conjugate base, the N1-deprotonated radical, G*(-H)*. Our ESR results clearly establish that at 77 K (i) one-electron oxidized guanine in double stranded DNA-oligomers exists as the deprotonated neutral radical G(-H)* as a result of facile proton transfer to the hydrogen bonded cytosine and (ii) the hole is preferentially located at the 5'-end in several ds DNA-oligomers with a GGG sequence.