Ionic liquids (ILs) are organic salts with low melting point by asymmetric ionic strength between cation and anion. They have been known as promising DNA extraction, separation and preservation agent due to their hydrophilic, hydrophobic interaction with DNA. However, few studies have been performed about how DNA-ILs complexes form and their mechanism. Herein, we present three types of ionic liquids (EMIM-Cl, BMIM-Cl, and OMIM-Cl) change the DNA structure depend on alkyl chain length of ionic liquids. Structural changes of DNA by ionic liquids are observed by Atomic force microscopy, gel electrophoresis, zeta potential and solid-state nanopore technology. The results of AFM show the different structures of DNA, including aggregate, stretching, and bundling shapes in terms of EMIM-Cl, BMIM-Cl, and OMIM-Cl respectively. In DNA translocation experiment, DNA/EMIM-Cl show rare translocation signal due to aggregated structure by neutralized surface charge. DNA/BMIM-Cl and DNA/OMIM-Cl show slowing down the translocation speed due to changes of DNA net charge and structure. Especially, OMIM-Cl make slowing down the DNA translocation speed about 102~104 times compared to translocation speed of bare DNA by unzipping the bundling shape of complex. In conclusion, the morphology of DNA could be modified by the incorporation with different alkyl chain length of ILs, providing their further application in nanopore technique for slowing DNA sequencing or understanding protein-DNA interaction.