Immobilized DNA is required for the development of DNA chips and arrays, DNA sensors, or other sensing devices including microfluidics, in addition to gene delivery devices. The broad application range for all of these DNA‐based systems is to a major extent found in the medical area, using the devices also in DNA sequencing and furthermore for food and environmental analyses. Therefore, strategies for DNA immobilization are described in this article, with focus on immobilization chemistry. Depending on the different surfaces, various immobilization techniques (e.g. via physical adsorption, covalent, affinity binding, and matrix entrapment) were developed and optimized, which are described for carbonaceous materials (e.g. carbon nanotubes), silica and silicon surfaces, gold surfaces, the same as for more recently complex biocompatible surfaces (e.g. polymeric gels). The ‘top‐down techniques’ can be distinguished from the ‘bottom‐up’ techniques started in the very beginning of DNA chip development. More recently, surfaces such as stents or biocompatible transplants are on focus to generate therapeutic devices. In the future, the power of DNA as a self‐assembling material could be used to combine both the ‘bottom‐up’ and ‘top‐down’ strategies to generate complex nanostructures with multifunctional applications (e.g. three‐dimensional DNA ‘origami’). Overall, the main aim of this article is to provide an overview of the methods currently used for DNA immobilization regarding the broad variety of the above mentioned potential applications.