Information security has become increasingly challenging as a result of massive advancements in information and communication technologies. Due to the necessity of exchanging private information and the open nature of the network, there is an increased risk of various types of attacks. Consequently, data security is an essential component of data communication. One of the most effective methods used to achieve secrecy is steganography. This method hides data within a cover object without raising suspicion. The level of security is improved when two steganography methods are combined. This approach is known as multilevel steganography, which hides sensitive data in two cover objects in order to provide a two-level security system. Accordingly, we developed a technique that focuses on protecting secrecy while also being robust to attacks. The new technique uses a multi-layer steganography mechanism by using DNA sequences and images as carriers for sensitive data. The technique intends to hide secret messages in the DNA using the substation algorithm, and then the fake DNA is embedded in an image utilizing the discrete cosine transform (DCT) method. Eventually, the stego image is sent to the intended recipient. Different types of images with different sizes and lengths of messages and DNA sequences were used during the experiments. The results show that the proposed mechanism is resistant to histogram and chi-square attacks. The maximum mean value observed was 0.05, which means the histograms of the original and stego images are nearly identical, and the stego image does not raise any suspicion regarding the existence of secret information. In addition, the imperceptibility ratios were good, as the highest PSNR and MSE values were 0.078 and 72.2, respectively. Finally, the PNG and BMP images show excellent results. On the other hand, the JPG images failed to meet the expected ratio of imperceptibility and security.