Miniaturized DNA sensitive field-effect transistors (DNA-FET) have been realized using single crystalline diamond grown by plasma-enhanced chemical vapor deposition (CVD). To bond DNA to diamond, amine linker-molecules are covalently attached by photochemical means to H-terminated diamond surfaces. Using hetero-bifunctional cross-linker and thiol-modified single-strand (ss) cancer marker DNA (CK20), the gate of diamond FETs is modified to sense hybridization of DNA, forming double-strand (ds) DNA molecules on the gate. The density of DNA bonded to diamond has been adjusted to about 1012 cm−2 and the experiments have been performed in phosphate buffer with different ionicity to control the Debye length of the Helmholtz layer. By hybridization, a gate-potential shift of 64 mV is detected in case of the 100 Å Debye lengths, while 46 mV is detected for 10 Å. This is discussed with respect to DNA related variations of charge and pH by hybridization. Time resolved experiments reveal exponential hybridization dynamics with a time constant of 600 s. The sensitivity limit of our experiment is about 1 nM.