Conventional methods for extraction of DNA are expensive, time-consuming and tedious. To overcome these limitations, a paper-based DNA extraction device was developed that incorporates sponge-based buffer storage, a paper-based valve and channels of different length to autonomously direct the reagent and sample to the Fusion 5 disk for DNA capturing. With this device, DNA can be extracted within 2 min from only 30 μL samples of whole blood, serum, breast cancer cell, saliva, sputum and bacterial suspension. The device can also extract Hepatitis B Virus DNA from clinical blood samples and after quantification shows a detection limit as low as 104 copies⋅mL−1. This highlights its potential use in future diagnostics. The performance of the device was similar to that of the commercial QIAamp DNA micro kits and the FTA card. In our perception, this simple, inexpensive, portable and disposable device holds great promise in terms of POC testing in resource-poor settings.