Pardee et al. described a novel approach to synthetic biology in which several engineered gene networks were freeze-dried onto paper with in vitro expression machinery and then rehydrated. The results showed high performance. These paper-based gene networks exhibited transformative potential for synthetic biology and its applications within the Department of Defense by shifting the power of engineered gene circuits from the fragile and challenging world of the cell onto stable, reproducible paper substrates. Paper-based gene networks offer a potential future for biodetection that is inexpensive disposable stable multiplexible over targets and modalities ribonucleic acidsmall moleculeprotein rapid to design and manufacture and embeddable in paper, clothes, and other porous materials. However, two major concerns challenged the potential of this brand new technology 1 whether its sensitivity could be augmented by gene-network amplification circuits and 2 whether cellular lysis could be embedded into the system, which would allow built-in sample preparation and maintain robust performance. In this study, we focused on the second concern and investigated whether paper-based gene networks exposed to bacterial lysate would function normally after being subjected to mechanical lysis.