In this review, we introduce a novel procedure to fabricate vertically aligned diamond nanowires with controlled geometrical properties like length and distance between wires by use of nanodiamond particles as a hard mask and by use of reactive ion etching. We summarize the characterizations of nanowires by atomic force microscopy and scanning tunneling microscopy as well as electrochemical techniques. In the last section, we show biofunctionalization of nucleic acid molecules on diamond nanowires using electrochemically bond nitrophenyl molecules as linker for deoxyribonucleic acid (DNA) sensing. The tip biofunctionalization and performance of as-prepared DNA sensors are discussed in detail.